Inhibiting mutant IDH-1

ABSTRACT

Methods of treating patients diagnosed with cancer harboring an IDH-1 mutation are provided, including the therapeutic administration of a certain inhibitor of a mutant IDH-1 as a single agent, or in combination with azacitidine (AZA).

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/414,505, filed May 16, 2019, which claims the benefit of and priorityto U.S. Provisional Application Serial Nos.: U.S. ProvisionalApplication No. 62/672,461, filed May 16, 2018; U.S. ProvisionalApplication No. 62/672,462, filed May 16, 2018; U.S. ProvisionalApplication No. 62/680,566 filed Jun. 4, 2018; U.S. ProvisionalApplication No. 62/680,571, filed Jun. 4, 2018; U.S. ProvisionalApplication No. 62/680,560, filed Jun. 4, 2018; U.S. ProvisionalApplication No. 62/680,562, filed Jun. 4, 2018; U.S. ProvisionalApplication No. 62/692,591, filed Jun. 29, 2018; U.S. ProvisionalApplication No. 62/692,598, filed Jun. 29, 2018; U.S. ProvisionalApplication No. 62/692,601, filed Jun. 29, 2018; U.S. ProvisionalApplication No. 62/692,604, filed Jun. 29, 2018; U.S. ProvisionalApplication No. 62/692,605, filed Jun. 29, 2018; U.S. ProvisionalApplication No. 62/773,562 filed Nov. 30, 2018; U.S. ProvisionalApplication No. 62/798,677, filed Jan. 30, 2019; U.S. ProvisionalApplication No. 62/798,681 filed Jan. 30, 2019; U.S. ProvisionalApplication No. 62/798,684, filed Jan. 30, 2019; U.S. ProvisionalApplication No. 62/798,687, filed Jan. 30, 2019; U.S. ProvisionalApplication No. 62/798,690, filed Jan. 30, 2019; and U.S. ProvisionalApplication No. 62/812,367, filed Mar. 1, 2019, the contents of each ofwhich are hereby incorporated herein by reference in their entirety.

TECHNICAL FIELD

The present disclosure relates to the medical treatment of certain formsof cancer harboring a mIDH1 mutation, and related medical conditions.

BACKGROUND

The metabolic enzyme isocitrate dehydrogenase (IDH1) catalyzes theoxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG). Inboth hematologic and solid tumor malignancies, mutated IDH1 acquires theneomorphic activity of converting α-KG to 2-hydroxyglutarate (2-HG) andthereby leads to the aberrant accumulation of 2-HG. 2-HG has beenproposed to act as an “oncometabolite” that has pleotropic effects ontumorigenesis. Excess production of 2-HG has been shown to inhibitα-KG-dependent enzymes involved in epigenetic regulation, collagensynthesis, and cell signaling, thereby leading to a block in normaldifferentiation of progenitor cells and the subsequent development ofcancer. Therefore, inhibition of mutated IDH1 in tumor cells and theconcomitant decrease in 2-HG production is a therapeutic approach to thetreatment of IDH1-mutated cancers.

IDH-1 mutations reported in cancer can occur at amino acid positionR132, such as R132H, R132C, R132S, R132G, and R132L mutations. Thereremains a need for therapeutic compounds that selectively inhibitproduction of 2-HG from cancer cells harboring a variety of R132 IDH-1mutations.

Indeed, mutant IDH-1 (“mIDH-1”) is a key therapeutic target for thetreatment of a variety of cancers and is the subject of extensiveresearch efforts to develop therapeutic compounds useful for theinhibition of mIDH-1. Many different small molecule inhibitors of m-IDH1have been disclosed in publications (e.g., WO2016/044789, WO2016/044787,WO2016/044782, WO2016/171755, and WO2016/171756). Among the other mIDH-1inhibitors that have been developed are those depicted in Table 1, whichincludes multiple compounds that have been reported in clinical trials,and certain other compounds that have been described as being useful forthe treatment of cancer.

TABLE 1

AG-120

AG-881

IDH305

IDH889

GSK321

Bay1436032

ZX-06

isocitrate dehydrogenase-1 inhibitor (Sanofi)

isocitrate dehydrogenase-1 inhibitor (Shanghai Haihe Pharmaceutical)

ABT-199

AG-221

The FDA recently approved the administration of 500 mg once daily of amutant IDH1 inhibitor for the treatment of acute myeloid leukemia (AML)with a susceptible IDH1 mutation as detected by an FDA-approved test in(a) adult patients with newly-diagnosed AML who are 75 years old or whohave comorbidities that preclude use of intensive induction chemotherapyand (b) adult patients with relapsed or refractory AML. However, a 2017Multi-Discipline Review panel at the FDA noted that doubling the drugdose of this compound translates to only approximately a 40% increase inexposure, and that an increase in clearance at steady state may berelated to autoinduction. A PBPK model reasonably captured theautoinduction effect of the 500 mg QD dose of this compound on CYP3A4,and described the steady-state PK profiles of this compound in patientsat clinically relevant exposure levels. The fold-change in relativebioavailability of this compound from single-dose to steady-state forthis compound was 0.50. In addition, following administration of asingle oral dose of 50 mg/kg to rats with an intact blood-brain barrier,this compound exhibited brain penetration of 4.1% (AUC_(0−8h)[brain]/AUC_(0−8h) [plasma]).

There remains an unmet medical need for a therapeutically effectivemethod of administering a mIDH-1 inhibitor providing the followingbenefits:

i) achieving sufficient predicted free brain drug exposure (e.g., adesirable predicted C_(brain) Ratio as disclosed herein) suitable fortreating mIDH1 solid tumors in the central nervous system, includingforms of brain cancer such as glioma; and

ii) achieving and sustaining a desired steady state drug plasmaconcentration within a suitable range for a patient throughout a desiredcourse of treatment (e.g., at least six months).

Thus, there remain particular challenges associated with treating m-IDH1forms of cancer, including the treatment of mIDH1 forms of blood cancer(e.g., AML) throughout a course of treatment (e.g., 15 days to 6months), treatment of mIDH1 forms of cancer across the blood brainbarrier (e.g., mIDH1 forms of glioma), and treatment of other mIDH1solid tumors.

SUMMARY

The administration of a therapeutically effective amount of Compound 1using the methods of treatment provided herein provides a treatment forpatients diagnosed with a cancer harboring a mIDH1 mutation, includingtreatment of such conditions of the blood (e.g., AML) and within thecentral nervous system (e.g., glioma). In some embodiments, a total of150 mg of Compound 1 can be administered twice per day on a daily basisto provide a therapeutically effective steady state blood concentrationthroughout a course of treatment (e.g., 6 months or more). This 150 mgBID dose of Compound 1 can be administered to a patient diagnosed with acancer harboring a mIDH1 mutation and continued each day throughout acourse of treatment. A course of treatment is preferably of sufficientduration to provide a therapeutically effective steady state plasmaconcentration in the patient diagnosed with the mIDH1 mutation (e.g., atleast 15 days), and continuing until a favorable treatment outcome isachieved. In some examples, a course of treatment is up to 6 months(e.g., 15 days to 6 months).

Compound 1 has been administered in human clinical trials as a singleagent or in combination with other therapeutic agents to treat multiplemIDH1 forms of cancer, including mIDH1 forms of AML and glioma. In oneexample provided herein, a patient diagnosed with mIDH1 AML treated with150 mg BID of Compound 1 achieved a complete remission after a 28 daycourse of treatment. In another example, a patient diagnosed with mIDH1AML was treated with 150 mg BID of Compound 1 in combination withazacitidine, as disclosed herein, for a total of eight 28-day treatmentcycles and also achieved complete remission of the AML. In a thirdexample, a patient diagnosed with mIDH1 glioma was treated with 150 mgBID of Compound 1 and demonstrated a partial response confirmed by MRI,after two consecutive 28-day treatment cycles, as determined by RANOcriteria (≥50% decrease in tumor, no new lesions, on stable dosecorticosteroids, no progression of measurable disease).

Compound 1 is a potent, selective, orally bioavailable, small-moleculeinhibitor of mutated IDH1 and is useful as an anticancer therapeutic inpatients with certain mutations in the IDH1 gene (including R132Xmutations).

mIDH1 forms of cancer can be treated by the therapeutic administrationof Compound 1 at a total daily dose of 300 mg (preferably, 150 mg twicedaily, or BID) to selectively inhibit multiple clinically relevant R132Xmutations of IDH-1. Compound 1 can be administered to achieve a durableeffective drug plasma concentration over the course of a desired courseof treatment (e.g., at least six months). Methods of administeringCompound 1 for the treatment of certain cancers, including glioma,hematological cancers such as AML, and solid tumors, are providedherein.

The present disclosure identifies a particular mIDH-1 inhibitor and,furthermore, new therapeutic methods for using this compound in thetreatment of cancer (e.g., AML, glioma, and various solid tumors). Inthe selection of the mIDH-1 inhibitor Compound 1, the present disclosureprovides an insight that certain prior assessments of multiple mIDH-1inhibitory compounds may have focused on and/or unduly prioritized oneor more features (e.g., in vitro potency as measured in biochemicalassays for the production of 2-HG) that can lead away from appreciationof certain unexpected properties of Compound 1 that lead to thediscovery of methods of treatment as described herein.

The discovery of the methods of treatment provided herein is based inpart on the selection of Compound 1 from among many compounds reportedto inhibit the production of 2-HG from mIDH-1 cancer cells. Compound 1was not initially reported as having the greatest biochemical potencycompared to reports for certain other small molecule inhibitors ofmIDH-1. Structurally distinct compounds (e.g., AG-120, AG-881, IDH305,IDH889, GSK321, and Bay1436032) and other certain quinolinone-basedcompounds were initially reported as having greater in vitro potency inbiochemical assays measuring activity against certain mIDH-1 isoforms.Compound 1 not only inhibits 2-HG production from cells harboringvarious R132X forms of IDH-1 mutations; Compound 1 is characterized by aCNS multiparameter optimization (“CNS MPO”) value supporting furtherdevelopment as an oral therapy for mIDH-1 forms of cancer in the centralnervous system.

Compound 1 has been used according to the novel methods of treatmentprovided herein to achieve both complete responses in the treatment ofmultiple patients diagnosed with AML in a human clinical trial, and apartial response in a human clinical subject continuing on therapy.Accordingly, the Applicants have discovered meaningful andtherapeutically beneficial methods of treating mIDH1 cancer that includethe selection of the mIDH1 inhibitor Compound 1 and the discovery ofcertain methods of administering Compound 1 that can provide steadystate blood plasma concentrations in a patient in need thereofthroughout a course of treatment that are suitable for treatment ofmultiple mIDH1 forms of cancer, including conditions of the blood (e.g.,AML) and the CNS (e.g., glioma).

The methods of treatment provided herein are based in part on multiplescientific discoveries, including: (1) the selection of Compound 1 ashaving sufficient in vitro potency against multiple R132X forms ofmIDH-1 cancer, (2) the properties of Compound 1 supporting its use inmIDH1 cancers in the central nervous system (e.g., predicted free braindrug levels of Compound 1 relative to the minimum effective level ofCompound 1, and subsequent partial response of a human mIDH1 gliomapatient to treatment with Compound 1), (3) the discovery of a specificdose and dose regimen for administering a therapeutically effectiveamount of Compound 1 useful for treating multiple mIDH1 cancers(including treatment of cancers of the blood, brain and other solidtumors) after comparative testing of multiple dose and dose intervals,and (4) the discovery that desired blood plasma concentration ofCompound 1 can be maintained in a patient throughout a course oftreatment of 6 months or more using the preferred methods ofadministering Compound 1 (e.g., 300 mg total per day, preferablyadministered as a total of 150 mg of Compound 1 BID).

IDH-1 Selectivity of Compound 1

Compound 1 selectively inhibits the production of 2-HG in mIDH-1 cancercells (i.e., cancer cells harboring IDH-1 R132X mutations) withdesirable in vitro potencies when compared to wild type IDH-1 cells andmIDH-2 cancer cells. In addition to this selectivity for mIDH-1,Compound 1 is active against multiple IDH-1 R132X mutants, and thereforecan be used to treat patients diagnosed with cancers possessing avariety of such mIDH1 mutations. Of these R132X IDH-1 mutants, R132H andR132C are the more frequently detected mutations for human mIDH-1cancers. With respect to inhibiting 2-HG production from mIDH-1 R132Cand R132H cell lines, Compound 1 shows comparable activity that iswithin 5-fold, compared to more disparate differences in activityranging from about 8-fold to 240-fold for comparative compounds [seeExample 4].

In addition to R132H and R132C IDH-1 mutants, Compound 1 inhibits theIDH-1 mutants R132L, R132G, and R132S. Notably, Compound 1 inhibits allfive of these IDH-1 mutants (R132L, R132G, R132S, R132H, and R132C) withonly a 7-fold range in potencies. Therefore, patients diagnosed withcancer possessing mutant IDH-1, e.g., having an IDH-1 R132X mutationselected from the group consisting of: R132L, R132G, and R132S (inaddition to R132H and R132C IDH-1 mutations), can be treated withCompound 1 (see Example 3).

Blood Brain Barrier Permeability of Compound 1

IDH-1 mutations in brain cancers, such as glioma, can result in abnormalhypermethylation of histones and DNA and suppression of normal cellulardifferentiation. IDH-1 R132H mutations represent more than 90% of theIDH mutations present in low grade glioma and secondary glioblastomamultiforme (GBM) patients. In addition, IDH-1 mutations R132C and R132Sare also reported in glioma patients. However, in order to be able totreat glioma, a small molecule inhibitor of mIDH-1 must be able to crossthe blood brain barrier (“BBB”) at a therapeutically effectiveconcentration over time, presenting another challenge to selecting acompound suitable for the treatment of glioma.

The ability to cross the BBB is by no means an intrinsic property ofmIDH-1 inhibitors. Referring to Table 13, many known mIDH-1 inhibitorshave undesirably low CNS MPO values (e.g., Table 13, compounds GSK321and Bay1436032). Therefore, even among these advanced drug candidates,there is no apparent reported correlation between mIDH-1 inhibitoryactivity and CNS MPO values.

In fact, selecting a compound that is both a potent inhibitor of mIDH-1and possesses a desirably high MPO value is not straightforward. Forexample, of the compounds specifically exemplified in WO/2016044789(“the '789 publication”), which include Compound 1, several compoundsare reported as having greater in vitro potency than Compound 1 in atleast one assay reported in Table 6 of the '789 publication (i.e.,compounds 1-20,1-22, 1-23, 1-25, 1-26, 1-27, and 1-29). See Table 13,which reproduces these data from the '789 publication. However, none ofthese compounds has a CNS MPO score as great as Compound 1. In fact, onecompound (1-26) does not even meet a desired minimum MPO threshold valueof 3.8 desired as a predictor of BBB permeability. Conversely, of thesix compounds with CNS MPO scores higher than Compound 1, five are lesspotent in vitro than Compound 1 in at least one biochemical assay of IDHinhibition (i.e., compounds 1-2, 1-3, 1-5, 1-6, and 1-11), with onecompound being “equipotent” (compound 1-1).

The lack of correlation between mIDH-1 inhibition and CNS MPO scores inTable 13 highlights the unpredictability in selecting a single mIDH-1inhibitor compound that inhibits multiple R132X forms of mIDH-1 withsufficiently similar potencies, and is also characterized by asufficiently high MPO score (e.g., 3.8 or higher), both of which aredesired in a therapy to treat a mIDH1 glioma. It is against thisbackdrop that the identification of Compound 1 as a compound having aMPO score of 3.8 or higher is unexpected. Indeed, rodent modeling showeda stark contrast between Compound 1 and two other reported mIDH-1inhibitors, AG-120 and AG-881. As described in Example 6, Compound 1partitions into the brain at a level 2-fold greater than that estimatedto achieve a therapeutic benefit, whereas AG-120 and AG-881 partitioninto the brain at a level less than what is estimated to achieve atherapeutic benefit. Thus, even when compared to another compound alsohaving a CNS MPO score suggestive of good BBB permeability, such asAG-881, these data indicate that Compound 1 possesses unexpectedlysuperior properties by combining desired comparative and selective invitro potency and predicted drug exposure in the brain.

For instance, as described in Example 6, preclinical studies show thatCompound 1 can cross the BBB in rodent models at desirable levels. Oraladministration of Compound 1 showed high systemic bioavailabilty inmultiple preclinical species. Permeability was excellent, with littleevidence of efflux, and significant brain penetration was observed inmice (98% brain binding in murine animal model). Based on theseassessments in rodents, Compound 1 is believed to cross the blood-brainbarrier to an extent effective to reach free concentration levels in thebrain consistent with pharmacological activity.

Dose and Administration of Compound 1

Upon identifying the unique characteristics of the mDH-1 inhibitorCompound 1 mentioned above, the next challenge was to determine asuitable dose of Compound 1 to be administered to human patients. Asuitable dose should possess an appropriate therapeutic index (e.g., anobserved in vivo efficacy against multiple forms of cancer harboringmIDH-1, but without unacceptable levels of toxic side effects). Morespecifically, a drug plasma concentration providing in vivo efficacy(“C_(eff)”) in patients with tumors producing 2-HG has been defined inthe literature as one that provides >90% inhibition of 2-HG production(Fan et al., 2014). Based upon mouse xenograft experiments described inExample 7 and plasma protein binding correlation in humans, Compound 1was found to have a C_(eff) of about 1,652 ng/mL. Therefore, preferredmethods of administering Compound 1 to treat a patient having a cancerharboring mIDH-1 can achieve a plasma concentration of at least about1,652 ng/mL.

In addition, preferred methods of administering Compound 1 avoidunacceptably high concentrations of Compound 1 in the patient. Thediscovery of the maximum preferred concentration of Compound 1 was basedin part on the results from a 28-day oral toxicity study in monkeys (seeExample 9), which found that the most significant adverse event was anincrease in mean QTc interval duration, a type of cardiac event that maycause arrhythmia. The lowest C_(max) plasma concentration of Compound 1at which prolonged QTc interval duration was observed was about 7,840ng/mL. Accordingly, Compound 1 is preferably dosed in a manner thatachieves a drug plasma concentration of no greater than about 7,800ng/mL (“C_(tox)”).

Preferred methods of administering a therapeutically effective amount ofCompound 1 provide a concentration of Compound 1 in the patient bloodplasma within a therapeutic range of about 1,652-7,840 ng/mL.

The discovery of provided methods of administering Compound 1 is basedon the evaluation of multiple doses of Compound 1 (100 mg, 150 mg and300 mg) at multiple dose intervals (once daily and twice daily), both asa single agent and in combination with another therapeutic agent.Notably, the administration of Compound 1 at a total daily dose of 300mg each day (preferably, 150 mg BID) ultimately demonstrated therapeuticbenefits in patient treatment across multiple forms of mIDH1 cancer. Asdescribed in Example 10, a Phase 1/2 study of Compound 1 was initiatedto evaluate Compound 1 alone or in combination with azacitidine (“AZA”)in mIDH-1 AML/myelodysplastic syndrome (MDS) patients. The blood plasmaconcentration and other effects of administering Compound 1 wereevaluated at multiple different dose of Compound 1 at different doseintervals: 100 mg QD (i.e., 100 mg once daily), 150 mg QD (i.e., 150 mgonce daily), 300 mg QD (i.e., 300 mg once daily), and 150 mg BID (i.e.,150 mg twice daily), with Compound 1 administered both as a single agentand/or in combination with AZA. As shown in FIG. 19 and described inExample 10, as a single agent, median C_(min) concentrations of Compound1 at steady-state were below C_(eff) in patients given doses of 100 mgor 150 mg once daily. When given at a dose of 300 mg once a day,Compound 1 achieved a median C_(min) above C_(eff), with a majority ofpatients reaching C_(eff). However, 150 mg given twice daily achievedthe highest median C_(min) and also showed less inter-quartilevariability in minimum concentrations as compared to 150 mg once daily.

Compound 1 was also administered in combination with azacitidine (AZA).As shown in FIG. 20, in this combination, dosing a total of 150 mg ofCompound 1 once daily shows a median C_(min) less than 1000 ng/mL, whichis well below C_(eff). In contrast, dosing a total of 150 mg of Compound1 twice daily shows a median C_(min) of over 3000 ng/mL, which is nearlytwice C_(eff).

Therefore, the present disclosure provides that a total daily dose of300 mg of Compound 1, preferably administered in a divided daily dose of150 mg BID, is preferred compared to other doses and dose intervalstested. Applicant selected 300 mg total daily dose, administered as a150 mg dose form. A suitable dose can be Compound 1 administered in a300 mg dose once daily. Preferably, a suitable dose can be Compound 1administered in a 150 mg dose twice daily.

Durable Steady State Concentration of Compound 1

Despite having found that a 300 mg total daily dose of Compound 1provides the desired efficacy, it was not clear whether this dosingwould be able to deliver a continued patient benefit for at least a sixmonth course of treatment (e.g., six four-week treatment cycles). Morespecifically, Applicant could not predict whether Compound 1 couldmaintain a steady state blood concentration (“C_(ss)”, as measured byC_(min) or trough concentration) without a substantial decline inconcentration over six four-week treatment cycles.

Compound 1 provides an unexpectedly durable steady state drug plasmaconcentration throughout a desired course of treatment. After theinitial 15 days of treatment with 150 mg twice daily of Compound 1, themedian steady state blood concentration of Compound 1 was maintainedabove about 2,000 ng/mL throughout a course of treatment (e.g., up toabout 36 weeks, including 12-32 weeks, as well as other intervalstherein, all measured from initial administration of Compound 1). Themedian C_(ss) was also well below the predicted threshold for QTcprolongation risk as discussed above. As shown in FIG. 8A, which showsdata for Compound 1 dosed as a single agent at 150 mg twice daily, themedian C_(ss) achieved on day 15 of cycle 1 (i.e., “C1D15, or on the15^(th) day of treatment with Compound 1) was maintained for at leastsix four-week treatment cycles, and in fact extended to beyond ninefour-week cycles. While concentrations are somewhat variable acrosspatients, the median C_(ss) level is maintained and does not trenddownward. Good retention of C_(ss) levels were also observed incombination with AZA as shown in FIG. 8B.

The discovery of the unique and remarkable properties of Compound 1 as aselective inhibitor of mIDH-1 across multiple mIDH-1 cancers led to thedevelopment of a dosing regimen of Compound 1 that overcomes therapeuticobstacles encountered with prior mIDH-1 inhibitor compounds. Through theadministration of Compound 1 at a total daily dose of 300 mg, thepresent disclosure provides sustained delivery of a mIDH-1 inhibitorat adesired drug plasma concentration for the treatment of cancer inpatients harboring mIDH-1.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A and FIG. 1B are a schematic of the dysregulation of epigeneticand gene expression profiles associated with IDH mutants.

FIG. 2 illustrates Compound 1 binding with mIDH.

FIG. 3A and FIG. 3B are each a schematic of diaphorase-coupled assaysused in Example 1, which measure activity by the determination of thelevel of remaining co-substrate NADPH after the enzymatic reaction isquenched.

FIG. 4A is a graph showing the results from a surface plasmon resonance(SPR) biophysical characterization of the molecular interaction betweenmIDH-1 inhibitor Compound 1 and recombinant IDH-1-R132H protein.

FIG. 4B is a comparator graph showing the SPR characterization ofCompound 1 at a BCL6 control surface.

FIG. 5 is a graph showing plasma and brain exposure for Compound 1following 5 mg/kg PO dose in a CD-1 Mouse.

FIG. 6 is a graph showing plasma and brain exposure for Compound 1following 100 mg/kg PO dose in a CD-1 Mouse.

FIG. 7A illustrates the summary of cohorts from a phase 1 study in mIDH1AML.

FIG. 7B illustrates use of Compound 1 in a phase 2 study in mIDH1 AMLand MDS.

FIG. 7C is a flow chart of human clinical trials of Compound 1, asdescribed in Example 10.

FIG. 8A is a graph of blood plasma concentration of Compound 1 measuredin a group of human patients treated with Compound 1 as a single agentthroughout a course of treatment. The dashed line labeled “C_(eff)” (islower than 1652 ng/mL.

FIG. 8B is a graph of blood plasma concentration of Compound 1 measuredin a group of human patients treated with Compound 1 and azacitidinethroughout a course of treatment. The dashed line labeled “C_(eff)” (islower than 1652 ng/mL.

FIG. 8C is a graph of effective blood plasma concentration of Compound 1measured in a group of human patients throughout a course of treatment.

FIG. 8D is a graph of blood plasma concentration of Compound 1 measuredin a group of human patients treated with Compound 1 and Azacitidinethroughout a course of treatment. The dashed line labeled “C_(eff)” islower than 1652 ng/mL.

FIG. 8E is a graph of blood plasma concentration of Compound 1 measuredin a group of human patients throughout a course of treatment.

FIG. 9A is a graph of the level of 2-HG in plasma of a group of humanpatients after consecutive treatment cycles of treatment with Compound 1as a single agent.

FIG. 9B is a graph of the level of 2-HG in plasma of a group of humanpatients after consecutive treatment cycles of treatment with Compound 1and azacitidine.

FIG. 9C is a graph of the level of 2-HG in plasma of a group of patientsafter two consecutive treatment cycles.

FIG. 9D is a graph showing the reduced level of 2-HG measured over timein a group of human patients treated with 150 mg of Compound 1 BID; thePD response is sustained throughout treatment with 150 mg of Compound 1BID.

FIG. 10A is a graph showing the levels of steady-state concentration ofCompound 1 measured in a group of human patients at the day 15 trough(ng/mL) of Compound 1 after administration to patients of 150 mg QD, 300mg QD, and 150 mg BID of Compound 1. FIG. 10A also illustrates steadystate is achieved by Week 2; (t_(1/2)˜60 hrs) and 150 mg BID steadystate exposures >IC₉₀ and below levels are expected to increase QTcFpotential.

FIG. 10B is a graph showing the 2-HG level measured in plasma (ng/mL) ina group of human patients after administration of 150 mg QD, 300 mg QD,and 150 mg BID of Compound 1.

FIG. 10C is a graph showing the plasma 2-HG reduction observed in ahuman patient population at all doses and schedules by cycle 2, day 1 ofadministration of Compound 1 and that a preferred PD response wasobserved with 150 mg BID of Compound 1.

FIG. 11A is a graph C_(ss) over time of patients treated with Compound 1at 150 mg BID. FIG. 11A also illustrates that plasma exposures ofCompound 1 are stable throughout treatment duration in patients. Thiscan serve as basis for selecting a 150 mg BID dose for dose expansionand a recommended phase II dose.

FIG. 11B is a graph plotting the ratio of steady state bloodconcentration of Compound 1 measured at different points during a Courseof Treatment in a human patient during the administration of Compound 1(150 mg BID). The Y-axis values are normalized to 1.0 using theconcentration measured on day 15 of a Course of Treatment. The data forthis graph were obtained from a single patient who received 150 mg BIDof the solid form of Compound 1 obtainable from Example 1 throughout aCourse of Treatment of over 300 days (i.e., greater than 6 months).

FIG. 12A is a graph showing the correlation between Compound 1 and 2-HGplasma levels in patients treated with Compound 1 as a single agentacross treatment groups and irrespective of time on treatment.

FIG. 12B is a graph showing the correlation between Compound 1 and 2-HGplasma levels in patients treated with Compound 1 and azacitidine incombination across treatment groups and irrespective of time ontreatment.

FIG. 13A, FIG. 13B, and FIG. 13C is a graph showing time on treatment ofAML patients treated with a single agent (Compound 1).

FIG. 14A and FIG. 14B is a graph showing time on treatment of AMLpatients treated with a combination of Compound 1 and azacitidine.

FIG. 15 is a graph showing cycle 1, grade 2 IDH-DS is resolved withdexamethasone and hydroxyurea. C2D1: CRp (1% blasts) to SCT in MLFS.

FIG. 16A is a graph of % patients who achieved transfusion independence(28- and 56-day) by category (CR/CRh, Non-CR/CRh, non-responders andoverall), illustrating transfusion independence by response category forpatients treated with Compound 1 as a single agent.

FIG. 16B is a graph showing blasts maximum reduction from baseline (%)based on category (CR/CRh, CRi, SD and PD), illustrating marrow blastreduction for patients treated with Compound 1 as a single agent.

FIG. 17A is a graph of % patients who achieved transfusion independence(28- and 56-day) by category (CR/CRh, Non-CR/CRh, non-responders andoverall), illustrating transfusion independence by response category forpatients treated with Compound 1 in combination with azacitidine.

FIG. 17B is a graph showing blasts maximum reduction from baseline (%)based on category (CR/CRh, CRi, SD and PD), illustrating marrow blastreduction for patients treated with Compound 1 in combination withazacitidine.

FIG. 18 consists of 4 panels: (a), (b), (c), and (d). Panel (a)illustrates free concentration of Compound 1 in plasma after three-doseoral administration (12.5, 25 and 50 mg/kg) with 12 hr dosing intervalin mouse HCT116-IDH1-R132H/+ xenograft model. Panel (b) illustrates freeconcentration of Compound 1 in tumor after three-dose oraladministration (12.5, 25 and 50 mg/kg) with 12 hr dosing interval inmouse HCT116-IDH1-R132H/+ xenograft model. Panel (c) illustrates percent2-HG inhibition in tumors in a PO dose of 12.5 mpk, 25 mpk and 50 mpk atthree different time points (4 h, 12 h, 24 h). Panel (d) illustrates invivo activity (2-HG % inhibition) of Compound 1 vs free compoundconcentration in tumor.

FIG. 19 is a graph of the minimum blood plasma concentration (Cmin) ofCompound 1 measured in groups of human patients receiving Compound 1 asa single agent at different dose amounts and dose intervals:day 15trough (ng/mL) of Compound 1 after administration to patients of 100 QD,150 mg QD, 300 mg QD, and 150 mg BID Compound 1 as a single agent. FIG.19 also illustrates steady state is achieved by Week 2; (t_(1/2)˜60 hrs)and 150 mg BID and 300 mg QD steady state exposures are >C_(eff).

FIG. 20 is a graph of the minimum blood plasma concentration (Cmin) ofCompound 1 measured in groups of human patients receiving Compound 1 atdifferent dose amounts and dose intervals, in combination withazacitidine: day 15 trough (ng/mL) of Compound 1 after administration topatients of 150 mg QD and 150 mg BID of Compound 1 in combination withazacitidine. FIG. 20 also illustrates steady state is achieved by Week2; (t_(1/2)˜60 hrs) and 150 mg BID QD steady state exposures are>C_(eff).

FIG. 21 illustrates a therapeutically reduced level of 2-HG in thepatient plasma after consecutive treatment cycles of treatment withCompound 1 as a single agent.

FIG. 22 illustrates a therapeutically reduced level of 2-HG in thepatient plasma after consecutive treatment cycles of treatment withCompound 1 and azacitidine.

FIG. 23 is a synthetic reaction scheme for the preparation of Compound1.

FIG. 24 depicts a dynamic vapor sorption (DVS) isotherm plot of Compound1 Type A solid form.

FIG. 25 depicts a differential scanning calorimetry (DSC) thermogram forCompound 1 Type A solid form.

FIG. 26 depicts a thermogravimetric analysis (TGA) curve for Compound 1Type A solid form.

FIG. 27 depicts X-ray powder diffraction (XRPD) of Compound 1 Type Asolid form.

DEFINITIONS

As used herein, the term “Course of Treatment” refers to the time periodin which a patient is being administered an agent, including anyadministration holidays or recovery periods. A course of treatment caninclude a single treatment cycle or multiple treatment cycles.Additionally, a course of treatment can include a partial treatmentcycle. The Course of Treatment can include the total time period duringwhich a patient is on a treatment protocol for a disease, e.g. AML orMDS, with a therapy comprising the administration of a mIDH-1 inhibitorcompound.

“Next-generation sequencing or NGS or NG sequencing” as used herein,refers to any sequencing method that determines the nucleotide sequenceof either individual nucleic acid molecules (e.g., in single moleculesequencing) or clonally expanded proxies for individual nucleic acidmolecules in a high-throughput fashion (e.g., greater than 103 or moremolecules are sequenced simultaneously). Various next generationsequencing methods are known. In one embodiment, the relative abundanceof the nucleic acid species in the library can be estimated by countingthe relative number of occurrences of their cognate sequences in thedata generated by the sequencing experiment. Next generation sequencingmethods are known in the art, and are described, e.g., in Metzker, M.(2010) Nature Biotechnology Reviews 11:31-46, incorporated herein byreference. Next generation sequencing can detect a variant present inless than 5% of the nucleic acids in a sample.

As used herein, the term “R132X mIDH-1 mutation(s)” refers to a mutationat the IDH-1 arginine 132 that results in inhibitory activity ofCompound 1 against the mutated IDH-1 form harboring the R132 mutation.Preferably, the R132X mutations have a 2-HG IC₅₀ value of less than 500nM (most preferably less than 250 nM or less than 150 nM) using the invitro assay of Example 2. Accordingly, preferred R132X mutations includeR132H and R132C, as well as R132L, R132G, and R132S (or other R132Xmutations having therapeutically relevant 2-HG IC₅₀ values obtainedusing the in vitro assay of Example 2). Patients having R132X mIDH-1mutation(s) can be identified using a suitable diagnostic, such as adiagnostic analyzing patient tissue with next generation sequencingtechnology that identified the presence of the R132X mIDH-1 mutation inthe patient tissue sample.

As used herein, the term “R132X mIDH-1 Selective Inhibitor Therapy”refers to a therapy administered to a patient to inhibit the activity ofR132X mIDH-1 in the patient, where the therapy is known to haveselective inhibitory activity against R132X mIDH-1 over wild type IDH-1.An R132X mIDH-a selective inhibitor therapy can be administration ofCompound 1 as disclosed herein.

As used herein, “sequencing” can be Next Generation Sequencing (NGS), ahigh-throughput sequencing technology that performs thousands ormillions of sequencing reactions in parallel. Although the different NGSplatforms use varying assay chemistries, they preferably generatesequence data from a large number of sequencing reactions runsimultaneously on a large number of templates. The sequence data can becollected using a scanner, and then assembled and analyzedbioinformatically. Thus, the sequencing reactions are performed, read,assembled, and analyzed in parallel.

The terms “subject” and “patient” are used interchangeably in thepresent disclosure.

Susceptible IDH1 mutations are defined as those leading to increasedlevels of 2-hydroxyglutarate (2-HG) in the specified mIDH1 cancer cells(e.g., mIDH1 leukemia cells or mIDH1 glioma cells) and where efficacy ispredicted by 1) clinically meaningful remissions with the recommendeddose of Compound 1 and/or 2) inhibition of mutant IDH1 enzymaticactivity at concentrations of Compound 1 sustainable at the recommendeddosage according to validated methods. Susceptible mutations includeR132H and R132C mIDH1 substitution mutations.

DETAILED DESCRIPTION

Compound 1 is a small molecule mIDH-1 inhibitor useful for the treatmentof patients harboring IDH-1 mutations. In some embodiments, Compound 1is a small molecule mIDH-1 inhibitor useful for the treatment of solidtumors in the CNS (e.g., glioma), hematological malignancies (e.g., AMLor MDS), or other solid tumors (e.g., chondrosarcoma, hepatobiliary, andintrahepatic cholangiocarcinoma).

Compound 1 can also be referred to as olutasidenib, CAS No. :1887014-12-1,(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile,5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile,or a compound of Formula (I).

Compound 1 has potent and equivalent biochemical activity against anumber of IDH-1 arginine 132 (R132) mutated forms, of which R132H andR132C are the most prevalent observed for human IDH-1. Compound 1 is asmall molecule mIDH-1 (mutated isocitrate dehydrogenase 1) inhibitor. Asshown in Example 3, it is a permeable, orally bioavailable compound,with an excellent preclinical profile in both in vitro and in vivomodels. Compound 1 is a potent and highly selective inhibitor of mutatedIDH1^(R132) enzymes:

TABLE 2 IDH1 Mutation R132H R132C R132G R132L R1325 IC₅₀ (nM) <10 <50<50 <10 <10

Compound 1 exhibits robust (>90%) knockdown of 2-HG in animal andneurosphere PD models. Compound 1 shows no CYP3A4 inhibition. Compound 1is blood brain barrier penetrant, showing total (bound) brain/plasmaratio of 30% in rats.

Compound 1 can be administered in combination with azacitidine. In someexamples, patients have been treated with or are already being treatedwith azacitidine. In some embodiments, a combination therapy of Compound1 and azacitidine can be administered for the treatment of patients witha cancer harboring IDH-1 mutations (e.g., mIDH1 forms of AML or gliomaor other solid tumors). For example, patients can be administeredCompound 1 daily (BID) in continuous 28-day cycles, in combination withazacitidine (administered at the dose of 75 mg/m² for 7 days IV/SC perevery 28-day cycle).

Isocitrate dehydrogenase (IDH) is a class of enzymes that catalyze theoxidative decarboxylation of isocitrate to α-keto-glutarate (α-KG).There are three isoforms in human cells. IDH-1 resides in the cytosoland peroxisomes, whereas IDH-2 and IDH-3 are mitochondrial enzymes.IDH-1 is dimeric and uses NADP+ as an electron acceptor. IDH-3 is atetrameric enzyme and, in contrast, uses NAD+ as an electron acceptor.IDH-3 is the primary IDH enzyme participating in the Krebs cycle. Thepresence of the IDH-1 mutations imparts a neomorphic activity to theenzyme, resulting in the production of (R)-2-hydroxyglutarate (2-HG)which has been termed an “oncometabolite”, and has pleotropic roles intumorigenesis.

Since IDH-1 mutations are only found in tumor tissue, the presentinvention is based in part on the discovery that the selective mIDH-1inhibitor of Compound 1 can be developed as a targeted therapy formultiple mIDH-1 forms of cancer. A patient selection biomarker for theuse of Compound 1 can be the existence of IDH-1 mutation in a patientdiagnosed with a cancer harboring mIDH-1. Studies in geneticallyengineered mouse models and models derived from cancer patient samplesboth support the discovery that mIDH produces 2-HG, the downstreameffects of which are believed to cause epigenetic changes that canconsequently block the proper differentiation of progenitor cells andlead to cancer. In particular, IDH-1 mutations can lead to the loss ofwild type enzymatic activity (conversion of isocitrate to α-KG).Instead, the mutated enzymes acquire the neomorphic activity ofconverting α-KG to 2-HG. In mIDH-1 harboring cancer cells, wild type andmutant IDH-1 form a heterodimeric complex that can produce very high2-HG levels. IDH-1 mutations can result in the formation of the(R)-enantiomer of 2-HG, which is in contrast to the accumulation of(S)-enantiomer found in L2HG aciduria patients, who harbor homozygousloss-of-function mutations in 2-HG dehydrogenase. Given the structuralsimilarity between 2-HG and α-KG, 2-HG has been shown to be acompetitive inhibitor of a number of α-KG dependent histone and DNAdemethylases. 2-HG can inhibit several KDM family histone demethylasesin vitro, including H3K9/H3K36 demethylases KDM4A and KDM4C, and H3K36demethylase KDM2A. Furthermore, elevated methylation levels of H3K4,H3K9, H3K27 and H3K79 have been observed in mIDH-1 containingpatient-derived samples, as well as in cells expressing IDH mutations ortreated with a cell-permeable ester of 2-HG. 2-HG can also inhibit theTET family of DNA demethylases, which in turn can result in thehypermethylation of DNA CpG islands. Mutations in IDH-1/2 and TET2 arethus far mutually exclusive, which supports the notion that 2-HGproduced by mIDH can inhibit TET2 and impair hematopoietic celldifferentiation. In addition, 2-HG has also been shown to block PHDactivity, which is critical for regulation of hypoxia inducible factorsand collagen hydroxylation and maturation. Hydroxylated collagen isimportant for the regulation of proliferation and proper differentiationof hematopoietic cells in bone marrow. Mutated IDH is also reported toblock proper hepatocyte differentiation and promote cholangiocarcinoma.

Using in vitro cellular mechanistic assays monitoring levels of theerrantly overproduced, tumorigenic metabolic byproduct 2-HG, inhibitionof mIDH-1 results in a >90% reduction in levels of measured 2-HG, aneffect that has also been shown to translate into similar levels of 2-HGsuppression in in vivo PK-PD studies in HCT116 (IDH-1 R132H) and HCT116(IDH-1 R132C) xenograft bearing mice. In both models, the freeconcentration of Compound 1 was comparable in plasma and xenografttumors, and exposures were dose dependent. At the highest dose tested inthese studies (50 mg/kg), Compound 1 inhibited 2-HG levels in tumorby >90% for up to 24 hours after the last dose in the HCT116 (IDH-1R132H) xenograft model, and to similar levels for at least 12 hours inthe HCT116 (IDH-1 R132C) model.

By far the most frequent IDH1 mutations occur at amino acid positionR132, and include R132H, R132C, R132S, R132G, and R132L mutations. Giventhat Compound 1 is a potent inhibitor of a spectrum of different IDH1R132 mutations, but is inactive against either wild type IDH1 or mutatedIDH2, patients will be selected based on the occurrence of an IDH1mutation at the R132 residue. Accordingly, Compound 1 is useful inmethods of treating patients diagnosed with a cancer harboring an IDH-1mutation such as, AML, MDS, glioma, hepatobiliary tumors,chondrosarcoma, cholangiocarcinoma or other solid tumors harboring anIDH1 mutation. For example, IDH-1 R132 mutations represent more than 90%of the IDH mutations present in low grade glioma and secondary GBMpatients. The neomorphic enzymatic activity acquired as a result ofIDH-1 mutation is believed to lead to the conversion of α-KG to 2-HG. Inconsequence, patients bearing IDH-1 mutations have elevated levels of2-HG. Most IDH-1 mutations result in a single amino acid change at theR132 residue, whereas most IDH-2 mutations occur at either Arginine 140(R140) or Arginine 172 (R172). The IDH mutation spectrum varies amongdifferent tumor types (Table 3).

TABLE 3 Total Tumor Mutation Types Frequency IDH Mutation IdentitiesGlioma 70-90% IDH1^(R132H), IDH1^(R132C), IDH1^(R132S), IDH2^(R172K) AML10-30% IDH2^(R140Q), IDH1^(R132H), IDH1^(R132C), IDH2^(R172K),IDH1^(R132G), IDH1^(R132S) Chondro- 75% IDH1^(R132C), IDH1^(R132H)sarcoma Intrahepatic 10-25% IDH1^(R132C), IDH1^(R132L), IDH1^(R132G),Cholangio- IDH1^(R132H), IDH2^(R172W) carcinoma

For example, IDH-1 R132 mutations represent more than 90% of the IDHmutations present in low grade glioma and secondary GBM patients. IDH-1mutations have been reported in hematological malignancies such as acutemyeloid leukemia (AML) and myelodysplastic syndrome (MDS), as well asmany solid tumors types, including low grade glioma, secondaryglioblastoma, intrahepatic cholangiocarcinoma (IHCC), chondrosarcoma,melanoma and hepatocellular carcinoma, specifically in cells which haveR132 mutation (e.g., R132C).

Compound 1 is an isocitrate dehydrogenase-1(IDH1) inhibitor useful forthe treatment of acute myeloid leukemia (AML) with a susceptible IDH1mutation as detected by an FDA-approved test. In some embodiments,pharmaceutical compositions comprising Compound 1 can be administered toadult patients with newly-diagnosed AML who are 75 years old or who havecomorbidities that preclude use of intensive induction chemotherapy. Insome embodiments, pharmaceutical compositions comprising Compound 1 canbe administered to adult patients with relapsed or refractory AML.

Patients can be selected for the treatment of AML with Compound 1 basedon the presence of IDH1 mutations in the blood or bone marrow. Patientswithout IDH1 mutations at diagnosis should be retested at relapsebecause a mutation in IDH1 may emerge during treatment and/or atrelapse. Information on FDA-approved tests for the detection of IDH1mutations in AML is available athttp://www.fda.gov/CompanionDiagnostics.

The patient can be diagnosed as having an IDH-1 R132 mutation disclosedherein using sequencing methods, such as next-generation sequencingmethods or a PCR based method. The diagnostic patient selection methodcan be a next-generation sequencing (NGS)-based tumor genotyping assayanalyzing a patient tissue sample, such as a bone marrow sample. Usefultechniques and technologies for diagnosing a patient as having a IDH-1R132 mutation may include, without limitation, sequencing machinesand/or strategies well known in the art, such as those developed byIllumina/Solexa (the Genome Analyzer; Bennett et al. (2005)Pharmacogenomics, 6:373-20 382), by Applied Biosystems, Inc. (the SOLiDSequencer; solid.appliedbiosystems.com), by Roche (e.g., the 454 GS FLXsequencer; Margulies et al. (2005) Nature, 437:376-380), and by others.

Next generation sequencing methods are known in the art.“Next-generation sequencing or NGS or NG sequencing” as used herein,refers to any sequencing method that determines the nucleotide sequenceof either individual nucleic acid molecules (e.g., in single moleculesequencing) or clonally expanded proxies for individual nucleic acidmolecules in a high-throughput fashion (e.g., greater than 103 or moremolecules are sequenced simultaneously). In one embodiment, the relativeabundance of the nucleic acid species in the library can be estimated bycounting the relative number of occurrences of their cognate sequencesin the data generated by the sequencing experiment. Next generationsequencing methods are known in the art, and are described, e.g., inMetzker, M. (2010) Nature Biotechnology Reviews 11:31-46, incorporatedherein by reference. Next generation sequencing can detect a variantpresent in less than 5% of the nucleic acids in a sample.

In some examples, patients treated with Compound 1 can have a mutantIDH-1 cancer that does not have a mIDH-2 mutation detected with a FDAapproved mIDH-2 diagnostic (e.g, as provided atwww.fda.gov/CompanionDiagnostics).

It will be appreciated that where the present disclosure refers to thetreatment of a patient, such disclosure includes the treatment of apopulation of patients as well (e.g., a population of patients having acancer harboring m-IDH1.

Pharmaceutical compositions comprising Compound 1 can be administeredthroughout a course of treatment. A course of treatment preferablycomprises administration of Compound 1 to a patient in need thereofuntil the concentration of Compound 1 in the blood of the patientreaches steady state (e.g., 15 consecutive days), until a desiredtherapeutic response (e.g., 6 months or more) or until diseaseprogression or unacceptable toxicity.

In some embodiments, Compound 1 can be formulated as a film-coated(e.g., 150 mg or 50 mg) tablet or capsule for oral administration. Eachtablet or capsule can contain the following inactive ingredients:colloidal silicon dioxide, croscarmellose sodium, hypromellose acetatesuccinate, magnesium stearate, microcrystalline cellulose, and sodiumlauryl sulfate. The tablet or capsule coating can include othercomponents such as FD&C blue #2, hypromellose, lactose monohydrate,titanium dioxide, and triacetin.

Compound 1 is a Selective mIDH1 Inhibitor

Compound 1 was selected as a potent and selective mIDH1 inhibitor. Thepresent disclosure provides methods for treating cancer. In particular,patients diagnosed with cancer harboring a mutant IDH-1 cancer cell,e.g., having a IDH-1 R132 mutation selected from the group consistingof: R132L, R132G, and R132S (in addition to R132H and R132C IDH-1mutations), can be treated with a therapeutically effective amount ofCompound 1. In some examples, patients treated with Compound 1 can havea mutant IDH-1 cancer that does not have a mIDH-2 mutation detected witha FDA approved mIDH-2 diagnostic (e.g., as provided atwww.fda.gov/CompanionDiagnostics).

Studies in genetically engineered mouse models and models derived fromcancer patient samples both support the discovery that mIDH produces2-HG, the downstream effects of which cause epigenetic changes thatconsequently block the proper differentiation of progenitor cells andlead to cancer. In particular, IDH-1 mutations can lead to the loss ofwild type enzymatic activity (conversion of isocitrate to alpha-KG(α-KG)). Instead, the mutated enzymes acquire the neomorphic activity ofconverting α-KG to 2-HG. In mIDH-1 harboring cancer cells, wild type andmutant IDH-1 form a heterodimeric complex that can produce very high2-HG levels. All IDH-1 mutations result in the formation of the(R)-enantiomer of 2-HG, which is contrast to the accumulation of(S)-enantiomer found in L2-HG aciduria patients, who harbor homozygousloss-of-function mutations in 2-HG dehydrogenase. Given the structuralsimilarity between 2-HG and α-KG, 2-HG has been shown to be acompetitive inhibitor of a number of α-KG dependent histone and DNAdemethylases. 2-HG inhibits several KDM family histone demethylases invitro, including H3K9/H3K36 demethylases KDM4A and KDM4C, and H3K36demethylase KDM2A. Furthermore, elevated methylation levels of H3K4,H3K9, H3K27, and H3K79 have been observed in mIDH-1 containingpatient-derived samples, as well as in cells expressing IDH mutations ortreated with a cell-permeable ester of 2-HG. 2-HG also inhibits the TETfamily of DNA demethylases, which in turn results in thehypermethylation of DNA CpG islands. Mutations in IDH-1/2 and TET2 arethus far mutually exclusive, which supports the notion that 2-HGproduced by mIDH inhibits TET2 and impairs hematopoietic celldifferentiation. In addition, 2-HG has also been shown to block PHDactivity, which is critical for regulation of hypoxia inducible factorsand collagen hydroxylation and maturation. Hydroxylated collagen isimportant for the regulation of proliferation and proper differentiationof hematopoietic cells in bone marrow. Mutated IDH is also reported toblock proper hepatocyte differentiation and promote cholangiocarcinoma.Since IDH-1 mutations are only found in tumor tissue, the presentinvention is based in part on the discovery of that the selective mIDH-1inhibitor of Compound 1 can be developed as a targeted therapy forcancer. The patient selection biomarker for the use of Compound 1 can bethe existence of IDH-1 mutation in a patient diagnosed with a cancerharboring mIDH-1.

Using in vitro cellular mechanistic assays monitoring levels of theerrantly overproduced, tumorigenic metabolic byproduct 2-hydroxyglutarate (2-HG), inhibition of mIDH-1 results in a >90% reduction inlevels of measured 2-HG, an effect that has also been shown to translateinto similar levels of 2-HG suppression in in vivo PK-PD studies inHCT116 (IDH-1 R132H) and HCT116 (IDH-1 R132C) xenograft bearing mice. Inboth models, the free concentration of Compound 1 was comparable inplasma and xenograft tumors, and exposures were dose dependent. At thehighest dose tested in these studies (50 mg/kg), Compound 1 inhibited2-HG levels in tumor by >90% for up to 24 hours after the last dose inthe HCT116 (IDH-1 R132H) xenograft model, and to similar levels for atleast 12 hours in the HCT116 (IDH-1 R132C) model.

Accordingly, Compound 1 is useful in methods of treating patientsdiagnosed with a cancer harboring an IDH-1 mutation. The neomorphicenzymatic activity acquired as a result of IDH-1 mutation is believed tolead to the conversion of α-ketoglutarate (alpha-KG) to2-hydroxyglutarate (2-HG). In consequence, patients bearing IDH-1mutations have elevated levels of 2-HG. Most IDH-1 mutations result in asingle amino acid change at the R132 residue, whereas most IDH-2mutations occur at either Arginine 140 (R140) or Arginine 172 (R172).The IDH mutation spectrum varies among different tumor types (Table 3above).

For example, IDH-1 R132 mutations represent more than 90% of the IDHmutations present in low grade glioma and secondary GBM patients. IDH-1mutations have been reported in hematological malignancies such as acutemyeloid leukemia (AML) and myelodysplastic syndrome (MDS), as well asmany solid tumors types, including low grade glioma, secondaryglioblastoma, intrahepatic cholangiocarcinoma (IHCC), chondrosarcoma,and melanoma. By far the most frequent IDH-1 mutations occur at aminoacid position R132, and include R132H, R132C, R132S, R132G, and R132Lmutations. Given that Compound 1 is a potent inhibitor of a spectrum ofdifferent IDH-1 R132 mutations, but is inactive against either wild typeIDH-1 or mutated IDH-2, patients will be selected based on theoccurrence of an IDH-1 mutation at the R132 residue.

The patient can be diagnosed as having an IDH-1 R132 mutation disclosedherein using sequencing methods, such as next-generation sequencingmethods. The diagnostic patient selection method can be anext-generation sequencing (NGS)-based tumor genotyping assay analyzinga patient tissue sample such as a bone marrow sample. Useful techniquesand technologies for diagnosing a patient as having a IDH-1 R132mutation may include, without limitation, sequencing machines and/orstrategies well known in the art, such as those developed byIllumina/Solexa (the Genome Analyzer; Bennett et al. (2005)Pharmacogenomics, 6:373-20 382), by Applied Biosystems, Inc. (the SOLiDSequencer; solid.appliedbiosystems.com), by Roche (e.g., the 454 GS FLXsequencer; Margulies et al. (2005) Nature, 437:376-380), and by others.

The invention is based in part on the discovery that Compound 1selectively inhibits the production of 2-HG from mIDH-1 cancer cellsharboring R132 mutations including R132S, R132G and R132L withclinically relevant comparative potencies, while remaining inactive atwild type IDH-1 cells. In addition, Applicants have discovered thatCompound 1 is a targeted, selective small molecule inhibitor of 2-HGproduction from mIDH-1 cancer cells and is also inactive in mIDH-2cancer cells that produce 2-HG (e.g., Compound 1 selectively inhibitsthe production of 2-HG from mIDH-1 cancer.

Solid Tumors

The invention is based in part on the discovery that Compound 1selectively inhibits the production of 2-HG from mIDH1 cancer cellsharboring R132 mutations including R132S, R132G and R132L withclinically relevant comparative potencies, while remaining inactive inwild type IDH1 cells. In addition, Applicants have discovered thatCompound 1 decreases 2-HG production in mIDH1 cancer cells, is aninhibitor of mutant IDH1 in cancer cells and is also inactive in mIDH2cancer cells that produce 2-HG (e.g., Compound 1 selectively inhibitsthe production of 2-HG from mIDH1 cancer).

In some examples, methods are provided for treating solid tumors in theCNS, including glioma cancer cells, harboring an IDH1 R132 mutation. Forexample, patients diagnosed with glioma harboring a mutant IDH1 cancercell can be treated with a therapeutically effective amount of Compound1 in combination with azacitidine.

Achieving Effective Blood Plasma Concentrations of Compound 1

The present disclosure provides methods for the treatment of AML or MDScomprising a step of administering to a subject a therapeuticallyeffective amount of a pharmaceutically acceptable form of Compound 1. Insome examples, the pharmaceutically acceptable form of Compound 1 is anoral dosage form (e.g., as provided in Example 1) administered to thepatient as R132X mIDH-1 Selective Inhibitor Therapy consisting of theoral administration of an oral dosage form of Compound 1 administeredeither as a single agent inhibitor of mIDH-1, or in combination withazacitidine or cytarabine. When Compound 1 is administered in suchcombination therapy, the subject can be receiving or have previouslyreceived treatment with azacitidine or cytarabine.

Post-therapy, 2-HG plasma levels less than 180 ng/mL are associated withbetter overall and disease-free survival in patients with IDH-1 andIDH-2 mutated AML (JANIN, M. et al., Serum 2-Hydroxyglutarate Productionin IDH1- and IDH2-Mutated De Novo Acute Myeloid Leukemia: A Study by theAcute Leukemia French Association Group, Journal of Clinical Oncology,32(4): 297-305 (2014)). FIG. 22 shows that 150 mg once daily dosingachieved a median 2-HG plasma concentration above 600 ng/mL. Meanwhile,300 mg once daily achieved a median 2-HG plasma concentration slightlyabove 200 ng/mL, and 150 mg twice daily achieved a median 2-GH HG plasmaconcentration of approximately 100 ng/mL. This demonstrates the abilityof 150 mg twice daily dosing to lower 2-HG levels below a literatureestablished level as compared to 150 mg and 300 mg once daily.

The invention is based in part on the discovery that oral administrationof Compound 1 (in the pharmaceutically acceptable oral dosage formresulting from the preparation method of Example 1) to humans havingelevated blood 2-HG levels (i.e., above about 180 ng/mL) can provide asteady state (trough) blood concentrations above a therapeuticallyeffective amount of Compound 1 (e.g., above the IC₉₀ concentration forR132H and/or R132C mIDH-1, and/or concentrations of greater than about2,000 ng/mL or concentrations of greater than about 1652 ng/mL)throughout a course of treatment of 6 months from the initialadministration of Compound 1, while simultaneously reducing and thenmaintaining the levels of 2-HG to below 200 ng/mL within about 15 daysof daily treatment with Compound 1. Alternatively, the levels of 2-HGare maintained below 180 ng/mL within about 15 days of daily treatmentwith Compound 1. In these observations, Compound 1 was administered inan oral dosage form (obtainable from the method in Example 1) twice perday (150 mg BID) to AML or MDS adult patients harboring a R132X IDH-1mutation. After the initial 15 days of treatment with 150 mg BID of thisoral dosage form of Compound 1, the steady state (trough) bloodconcentration of Compound 1 (pre-dose) was maintained above about 2,000ng/mL (and well below the predicted threshold for QTc risk) throughout acourse of treatment (e.g., up to about 30 weeks, including 12-30 weeks,and 20 weeks as well as other intervals therein, all measured frominitial administration of Compound 1).

Accordingly, the present disclosure provides methods of treatingpatients harboring isocitrate dehydrogenase 1 mutations (mIDH-1)(preferably including one or more R132X mIDH-1) diagnosed with AML orMDS. The method can comprise administering to the patient in needthereof a therapeutically effective amount of a R132X mIDH-1 SelectiveInhibitor Therapy. The R132X mIDH-1 selective inhibitor can consist ofCompound 1 as the only R132X mIDH-1 inhibitor compound administered tothe patient (e.g., in an oral dosage form such as the solid formobtained from Example 1). Compound 1 can be administered to a patientharboring the R132X mIDH-1 identified in a tissue sample, and/or anelevated 2-HG blood concentration (e.g., above about 180 ng/mL) for acourse of treatment of at least three consecutive treatment cycles of 28consecutive days of administration for each cycle. The course oftreatment can start with the initial administration of Compound 1 in thefirst of the at least three or more consecutive 28-day treatment cycles.The administration of the therapeutically effective amount of Compound 1throughout a course of treatment (e.g., at least 15 consecutive days,preferably up to 30 weeks or more) to a patient having elevated 2-HGlevels (e.g., 2-HG blood concentrations in plasma of 200-10,000 ng/mL)can result in a therapeutic effect on the patient evidenced by a durabletherapeutically effective trough blood plasma concentration of Compound1 in the patient throughout the course of treatment (e.g, above the IC₉₀concentration for R132H and/or R132C mIDH-1, and/or concentrations ofgreater than about 2,000 ng/mL and less than about 7,200 ng/mL, or abovethe IC₉₀ concentration for R132H and/or R132C mIDH-1, and/orconcentrations of greater than about 1652 ng/mL and less than about7,840 ng/mL).

Compound 1 can be administered at a dose of 150 mg twice per daythroughout the course of treatment. Compound 1 can be administered withfood to improve bioavailability of Compound 1. The course of treatmentcan be at least 15 consecutive days starting with the initial dose ofCompound 1 and longer (e.g., up to 30 weeks, 15 days to 30 weeks, 15days to 12 weeks, at least 12 weeks, 12-30 weeks, 15 days to 6 monthsand other intermediate or longer durations or intervals apparent basedon the present disclosure).

Some methods further comprise the administration of azacitidine to thepatient throughout the course of treatment. Azacitidine can besubcutaneously or intravenously administered to the patient in anazacitidine treatment cycle consisting of the administration of a totaldose of 75 mg/m² each day for 7 consecutive days beginning at the startof each treatment cycle, followed by 21 consecutive days withoutadministration of the azacitidine to the patient. A 48-hourdose-interruption of azacitidine is allowed for weekends or holidays. Ifno response is seen after 2 treatment cycles, Azacitidine can beadministered at a total dose of 100 mg/m² each day. Treatment with IDH1minhibitor and azacitidine showed synergistic effects on releasingdifferentiation block in mIDH leukemia models in vitro.

The methods can further comprise the administration of cytarabine to thepatient throughout the course of treatment. Cytarabine can besubcutaneously or intravenously administered to the patient in acytarabine treatment cycle consisting of the administration of a totaldose of 20 mg/day each day for 7 consecutive days beginning at the startof each treatment cycle, followed by 10 consecutive days withoutadministration of the cytarabine to the patient. Cytarabine can also beadministered 20 mg BID subcutaneously for 10 days beginning at the startof each treatment cycle. In the induction therapy of AML, the cytarabinedose administered in combination with other anticancer drugs can be 100mg/m²/day by continuous IV infusion (Days 1 to 7) or 100 mg/m² IV every12 hours (Days 1 to 7). Cytarabine injection can be used intrathecallyin acute leukemia in doses ranging from 5 mg/m2 to 75 mg/m² of bodysurface area. The frequency of administration can vary from once a dayfor 4 days to once every 4 days. The dose can be 30 mg/m² every 4 daysuntil cerebrospinal fluid findings are normal, followed by oneadditional treatment.

A patient can be identified as having a R132X mutation in mIDH-1 using adiagnostic method comprising a sequencing analysis (e.g., nextgeneration sequencing (NGS)) of bone marrow or other tissue sampleobtained from the patient prior to the administration of Compound 1 tothe patient. The R132X gene mutation can be determined prior toadministration of Compound 1 to the patient. Compound 1 can beadministered to patients who have received prior anticancer therapyand/or other concomitant (non-anticancer) medications. In some examples,Compound 1 is administered to patient who has not received a priormIDH-1 inhibitor therapy.

As provided herein, methods for the administration of an inhibitor ofthe R132X mutant IDH-1 (mIDH-1 Inhibitor) Compound 1 provide anunexpectedly durable steady state blood concentration of the mIDH-1Inhibitor throughout a desired course of treatment. For example, thetherapeutic methods provided herein can provide AML or MDS patientsharboring a R132X mIDH-1 mutation with durable steady state bloodconcentrations of the mIDH-1 Inhibitor of Compound 1 at atherapeutically effective level (e.g., above the IC₉₀ concentration fora R312X mIDH-1) without a substantial decline (e.g., no more than 10%reduction) in initial Compound 1 mIDH-1 Inhibitor steady state bloodconcentration (e.g., blood concentration measured about 12 hours afteran initial dose of the mIDH-1 Inhibitor) over a course of treatment ofgreater than about 12 consecutive weeks (e.g., 3 consecutive 28-daytreatment cycles and preferably at least about 6 months). In addition,the mIDH-1 Inhibitor Compound 1 can be administered to AML or MDSpatients harboring a R132X mIDH-1 mutation in a therapeuticallyeffective manner that provides for the reduction of elevated 2-HG levelswithin about 15 days of initiating a course of treatment, preferablyachieving and maintaining 2-HG levels in these patients at a level at orbelow about 180 ng/mL starting by day 15 in a course of treatment andcontinuing throughout a course of treatment lasting for at least 12weeks or longer (e.g, 12-30 weeks).

Referring to FIGS. 8A-8E the administration of Compound 1 in the oraldosage form described in Example 1 at 150 mg BID resulted in a sustainedtherapeutically effective trough blood plasma concentration above 2,000ng/mL after cycle 3 of a 28-day treatment cycle. Simultaneously,referring to FIGS. 9A-9D, the administration of Compound 1 in the oraldosage form from Example 1 at 150 mg BID resulted in a sustained 2-HGlevel (e.g., under 200 ng/mL in plasma after cycle 3 day 1 of a 28-daytreatment cycle). This effect is in contrast to other IDH1 inhibitorsthat have been advanced clinically. Accordingly, the invention is alsobased in part on the discovery that administration of Compound 1 at 150mg BID results in a sustained ratio of greater than about 10 (preferablygreater than about 20) of blood plasma concentration of Compound 1(e.g., trough concentrations measured pre-dose at concentrations ofabout 2,000 ng/mL or greater) to 2-HG blood level (e.g, plasmaconcentrations of about 200 ng/mL or lower, including concentrations ofabout 100 ng/mL) after cycle 3 day 1 (i.e., after BID doses administeredover the initial 15 consecutive days of treatment) of a 28-day treatmentcycle. A plasma half-life of about 60 hours was estimated for Compound1, with steady state achieved by week 2 of the course of treatment. Thesteady state blood concentrations of Compound 1 measured in the patientswas above the IC₉₀ value for 2-HG inhibition in R132X mIDH-1 cells(described in the Examples). As shown in FIGS. 8A-8E, the plasmaexposures (steady state blood plasma concentration) of Compound 1 weredurable (i.e., sustained) throughout the 30-week treatment duration. Asshown in FIGS. 9A-9D the plasma 2-HG concentrations were reduced to thenormal range within 1 cycle (C2D1) and maintained throughout thetreatment duration. No dose limiting toxicities of Compound 1 wereobserved during dose escalation studies, and the maximum tolerated dose(MTD) of Compound 1 was not reached. FIGS. 8A-8E are graphs of the dataobtained from measuring the steady state concentrations from patients inthe clinical trial receiving a 150 mg BID dose of Compound 1, either asa single agent or in combination with azacitidine as described inExample 10. As shown in FIG. 8D, azacitidine did not alter thepharmacokinetics of Compound 1, with consistent Compound 1 plasmaconcentrations observed over the treatment duration. FIG. 11A is a graphshowing the steady state concentration of Compound 1 measured inpatients at various points during the Course of Treatment described inthe clinical trial of Example 10, with each point representing a cyclenumber and day number (each cycle is 28 consecutive days ofadministration of 150 mg BID Compound 1). The steady state concentrationof Compound 1 remained above the minimum desired level (bottom dashedline) and the maximum desired level (upper dashed line).

Compound 1, with a plasma half-life of ˜60 hours, achieved steady-stateconcentration within 2 weeks of dosing and remained consistent overtreatment duration. At 150 mg BID (RP2D) as a single agent, steady-stateplasma concentrations are above the preclinical C_(eff) resulting in 90%reduction in plasma 2-HG and below Compound 1 levels predicted, in NHP,to pose a QTc prolongation risk. At 150 mg BID as a single agent, asignificant reduction (p<0.0009) in plasma 2-HG levels was achieved byend of Cycle 1 and was sustained over the treatment duration.Combination therapy of azacitidine plus Compound 1 150 mg BID achievedsteady state concentrations of Compound 1 above the preclinical C_(eff),resulting in ≥90% reduction in plasma 2-HG and below Compound 1 levelspredicted to pose a QTc prolongation risk. A significant (p<0.0001)reduction in plasma 2-HG levels at the end of Cycle 1 was observed withthe combination of Compound 1 (150 mg BID) and azacitidine and wassustained over the treatment duration. However, a slower rate of declinecompared to the single agent Compound 1 has been observed.

The PK/PD relationship of individual subjects' plasma Compound 1 and2-HG concentration across single agent treatment groups and irrespectiveof time on treatment is presented in FIG. 12A. Compound 1 concentrations<1,000 ng/mL correspond to early time point assessments (C1D0-C1D15). Nocorrelation of exposure to 2-HG reduction is observed until Compound 1concentrations crossed the C_(eff) predicted by in vivo models to resultin >90% reduction in plasma 2-HG levels. As shown in FIG. 12A, nocorrelation of exposure to 2-HG reduction is observed until Compound 1concentrations crossed the C_(eff) predicted by in vivo models to resultin >90% reduction in plasma 2-HG levels. Maximum, most consistentexposure-response was observed at the median C_(ss) of Compound 1 with150 mg BID (RP2D).

The PK/PD relationship of individual subjects' plasma Compound 1 and2-HG concentration across combination of Compound 1 and azacitidinetreatment groups and irrespective of time on treatment is presented inFIG. 12B. Compound 1 concentrations <1,000 ng/mL correspond to earlytime point assessments (C1D0-C1D15). No correlation of exposure to 2-HGreduction is observed until Compound 1 concentrations crossed theC_(eff) predicted by in vivo models to result in >90% reduction inplasma 2-HG levels. Maximum, most consistent exposure-response wasobserved at the median C_(ss) of Compound 1 with 150 mg BID (RP2D).

FIGS. 13A and 13C are graphs showing the results from the clinical trialin Example 10, showing the 150 mg BID administration Compound 1 (solidform obtainable from Example 1) was administered as a single agent tomultiple patients over a Course of Treatment with times having a medianof 94 days, and a range of 1 to 586 days. About 32% of the patientsremain on treatment.

FIG. 13B depicts responses for relapsed or refractory (R/R) AMLpatients, and shows prolonged duration of treatment with Compound 1 wasobserved with a 1st response occurring within 2 months of treatment withCompound 1. Responses <CR/CRi were noted to deepen with continuedtreatment resulting in CR/CRh/CRi rate of 41% in R/R AML. Clinicalbenefit (SD ≥8 weeks) was observed in subjects without an IWG definedresponse. 10% of patients (1 AML and 2 MDS) remained on treatment.Treatment Discontinuation: Progressive Disease (PD) (9), death (6),transplant (4), Adverse Events (AE) (3), investigator's decision (2),withdrawal of consent (1), and lack of response (3).

Summary of Response for Compound 1 as a Single Agent

TABLE 4 R/R AML All AML & MDS Response N = 22 N = 31 ORR, n (%)* 9 (41)11 (35)  [95% Cl] [21, 64] [19, 55] CR, n (%) 4 (18) 5 (16) CRh, n (%) 2(9)  2 (6)  CRi, n (%) 3 (14) 4 (13) SD, n (%) 5 (23) 11 (35)  PD, n (%)1 (5)  1 (3)  NE, n (%) 7 (32) 8 (26) *ORR = overall response rate: CR +CRh + CRi + PR + MLFS. No PR/MLFS observed.

Administration of Compound 1 at 150 mg BID in the clinical trial ofExample 10 reduced the measured levels of 2-HG in the blood of patientsas shown in FIGS. 9A-9D and FIGS. 10B-10C. The 2-HG levels measured inthe blood of patients during this clinical trial of Example 10 are shownin FIGS. 9A-9D, demonstrating reduction of 2-HG levels within about 1-228-day treatment cycles (150 mg Compound 1 is administered BID on eachday for 28 consecutive days for each treatment cycle, as described inExample 10).

FIGS. 10B-10C are each a graph showing the concentration of 2-HGmeasured in the blood of patients receiving one of three different doseand dose intervals: 150 mg QD, 300 mg QD or 150 mg BID (either receivingCompound 1 as a single agent or in combination with azacitidine asdescribed in the clinical trial of Example 10, in each category). The2-HG levels are measured prior to administration of Compound 1, and thenmeasured after administration of Compound 1 up to cycle 2, day 1 afterfirst receiving Compound 1 (as the solid form obtained from Example 1).

The invention is based in part on the discovery that administration ofCompound 1 at 150 mg BID resulted in a higher blood exposure level thaneither 150 mg QD or 300 mg BID at day 15. See, for example, FIG. 10A.Administration of Compound 1 at 150 mg QD BID resulted in a bloodexposure level of <3000 ng/mL at day 15. Administration of Compound 1 at300 mg QD BID did not result in improved blood levels at day 15. Incontrast, administration of Compound 1 at 150 mg BID results in a bloodexposure level of >3000 ng/mL at day 15. Additionally, the invention isbased in part on the discovery that administration of Compound 1 at 150mg BID resulted in a lower 2-HG level in plasma than either 150 mg QD or300 mg BID at day 15. See, for example, FIG. 10B.

The oral dosage form of Compound 1 (Example 1) was administered to humanpatients as a single agent (150 mg QD, 300 mg QD, 150 mg BID and 100 mgQD until disease progression) in a human clinical trial treating AML/MDSin cancer patients harboring a mIDH1 mutation, as described in theExamples below. FIG. 10A is a graph showing the concentration ofCompound 1 measured in the blood of patients after receiving Compound 1(as the solid form obtained from Example 1) in one of three differentdose and dose intervals: 150 mg QD, 300 mg QD or 150 mg BID (eitherreceiving Compound 1 as a single agent or in combination withazacitidine as described in the clinical trial of Example 10, in eachcategory).

The present disclosure includes methods for treating AML or MDS inpatients having one or more R132X mIDH-1 mutations (e.g., as measured ina tissue sample obtained from the patient) and/or elevated 2-HG levels(e.g., 2-HG levels measured in a blood sample at above about 180 ng/mL),comprising administration of Compound 1 alone (e.g., as a single agent)or in combination with azacitidine or cytarabine. For methods whereCompound 1 is administered as a combination, the subject beingadministered Compound 1 may be receiving or previously receivedtreatment with azacitidine or cytarabine.

The methods of treatment can include the administration of Compound 1such that on day 1 of cycle 4 of repeated 28-day treatment cycles (orday 1 of any subsequent cycle), the trough blood plasma concentration ofCompound 1 has not decreased more than about 5-25%, about 5-20%, about5-15%, about 5-10%, about 10-25%, about 10-20%, or about 10-15%, ascompared to the trough blood plasma concentration on day 1 of cycle 2.Preferably, patients harboring a R132X mIDH-1 mutation can beadministered 150 mg of Compound 1 twice daily (BID) every day onconsecutive days (without holiday) for one or more continuous 28-daycycles.

Compound 1 can be administered to certain patients in combination with ahypomethylating agent such as azacitidine. IDH1 mutations (e.g., in AMLor MDS patients harboring a R132X mIDH-1 mutation) can result inabnormal hypermethylation of histones and DNA and suppression of normalcellular differentiation. The combination of Compound 1 and azacitidinecan be administered for the treatment of patients with AML harboringIDH1 mutations. For example, patients can be administered the Compound 1daily (BID) in continuous 28-day cycles, alone or in combination withazacitidine (administered at the dose of 75 mg/m² for 7 days IV/SC perevery 28-day cycle). For example, Compound 1 can be administered at adose of 150 mg QD or 150 mg BID in combination with azacitidine(azacitidine administered per standard of care for a patient). FIG. 14is a graph showing the results from the clinical trial in Example 10,showing the 150 mg BID administration Compound 1 (solid form obtainablefrom Example 1) in combination with the administration of azacitidine(administered at the dose of 75 mg/m² for 7 days IV/SC per every 28-daycycle), administered to multiple patients over a Course of Treatmentwith times having a median of 87 days, and a range of 10 to 351 days.About 48% of the patients remain on treatment.

As shown in FIG. 13B, upon further duration of treatment, clinicalresponses observed in R/R or TN AML and MDS: cytopenias associated withazacitidine may influence depth of IWG response. An ORR of 46% wasobserved for the combination of Compound 1 with azacitidine in R/R AMLand an ORR of 78% was observed in TN AML (CR/CRi of 66%). TreatmentDiscontinuation was caused by: PD (6), transplant (6), investigator'sdecision (5), death (4), AE (2), and others: treatment failure, hospice,other treatment (1 each). 37% of patients (AML and MDS) remain ontreatment.

TABLE 5 Summary of Response for Compound 1 as a single agent R/R AML TNAML All AML & MDS Response N = 26 N = 9 N = 41 ORR, n (%)* 12 (46) 7(78)  22 (54) [95% Cl] [27, 67] [40, 97] [37, 69] CR, n (%)  3 (12) 2(22)  8 (20) CRh, n (%) 1 (4) 0  1 (2) CRi, n (%)  5 (19) 4 (44)  9 (22)PR, n (%) 1 (4) 1 (11) 2 (5) MLFS, n (%) 2 (8) 0 2 (5) SD, n (%) 11 (42)1 (11) 14 (34) PD, n (%) 1 (4) 0 1 (2) NE, n (%) 2 (8) 1 (11)  4 (10)*ORR = CR + CRh + Cri + PR + MLFS

In some methods, Compound 1 can be administered with cytarabine. Lowdose cytarabine (LDAC) can be administered to certain AML patients(e.g., AML patients at or above about 60 years of age who are notcandidates for intensive therapy, and harboring a R132X mIDH-1mutation). The therapeutically effective combination of Compound 1 withLDAC can be administered to AML patients harboring IDH1 mutation. Forexample, patients can be administered the Compound 1 daily (BID) incontinuous 28-day cycles, alone or in combination with LDAC(administered at the dose of 20 mg BID SC for 10 days every 28-daycycle) until treatment discontinuation.

Subjects that are treated according to provided methods and combinationtherapies can have relapsed or refractory AML or MDS, or “high risk”MDS, and may have been previously treated with a mIDH1 inhibitor. Suchrefractory AML or MDS (i) can be naïve to prior hypomethylating therapyand IDH1 inhibitor therapy and/or (ii) may have shown inadequateresponse or progressed immediately preceding hypomethylating therapy.The provided methods and combination therapies can be used to treatsubjects with residual IDH-R132 mutations. The provided methods andcombination therapies can also be used to treat subjects with AML or MDSin morphologic complete remission or complete remission with incompleteblood count recovery (CR/CRi) after cytotoxic-containing inductiontherapy.

The methods of treatment are based in part on a human clinical study ofadministration of Compound 1 in 3 stages: a phase 1 dose-escalationstage, a phase 1 dose-expansion stage and a phase 2 stage, as furtherdescribed in the Examples. Single agent Compound 1 dose escalation wasadministered in once-daily (QD) doses of 150 and 300 mg, a twice-daily(BID) dose of 150 mg or a once daily dose of 100 and 150 mg with food topotentially improve bioavailability. During the course of single agentdose escalation, a parallel escalation arm can be initiated for Compound1 in combination with azacitidine. This combination can be initiatedonce the first dose level cohort of Compound 1 in the single agentschedule (150 mg QD) is complete. Once the maximum tolerated dose or themaximum evaluated dose is identified for the single-agent andcombination cohorts, select populations of patients can be enrolled intophase 1 dose expansion cohorts at the selected single agent orcombination doses, to further characterize the safety profile andconfirm the recommended phase 2 dose. After the recommended phase 2 dosein combination with azacitidine is selected, a cohort of 6 patients aretreated with Compound 1, at that dose, in combination with low dosecytarabine. In the Phase 2 portion, specific populations of patientswith AML/MDS harboring IDH1-R132 mutations are enrolled to receiveCompound 1 either as a single agent or in combination with azacitidineat the recommended phase 2 doses.

As outlined in Examples 10 and 12, Compound 1 demonstrates clinicalactivity as single agent in a high-risk Phase 1 population of AML/MDSpatients with IDH1 mutation. 41% CR/CRh/CRi in R/R AML (35% in allAML/MDS) was observed in patients treated with Compound 1 as a singleagent. Transfusion independence was observed in both IWGresponders/non-responders. Durable disease control or stable disease4-12+ months was observed in R/R AML. An observed reduction in bonemarrow blasts is supportive of clinical benefit to patients. Compound 1was well tolerated with patients maintained in treatment for a median of5.6 months, likely contributing to rate and depth (CR/CRh) of response.Compound 1 plasma exposure correlates with 2-HG response. Compound 1C_(ss) reduction of 2-HG supports 150 mg BID as the dose and scheduleselected for evaluation in global Phase 2 trial outlined in FIG. 7B.

As outlined in Examples 10 and 12, the combination of Compound 1 andazacitidine demonstrates clinical activity in a high-risk Phase 1population of AML/MDS patients with IDH1 mutation. Patients maintainedtreatment for a median of 5 months. Durable disease control (>6 months)was observed in the absence of IWG response. 46% ORR and 35% CR/CRh/CRiin R/R AML, 78% ORR in TN AML was observed. Compound 1 is well toleratedin combination with azacitidine and possesses low risk of QTprolongation (2 AEs reported). Azacitidine combination modestlyincreased metabolic and gastrointestinal treatment emergent adverseevents (TEAEs). Higher rates of neutropenia compared to SA treatment(Grade 3/4 17% vs 6%) were observed which may be impacting the depth(CR/CRh) response. Compound 1 plasma exposure was shown to correlatewith 2-HG response. Compound 1 C_(ss) reduction of 2-HG supportsselection of 150 mg BID as RP2D.

In some embodiments, the present disclosure additionally providesmethods of treating AML or MDS in a patient harboring isocitratedehydrogenase 1 mutations (mIDH1), which can comprise administering to apatient in need thereof a therapeutically effective amount of Compound 1each day for at least three consecutive treatment cycles of 28consecutive days each. The administration of a therapeutically effectiveamount of Compound 1 can result in the patient having a durabletherapeutically effective trough blood plasma concentration of Compound1 in the patient throughout the course of treatment.

In some embodiments, the administration of a therapeutically effectiveamount of Compound 1 can result in the level of 2-HG in the patient'splasma being maintained at or below about 200 ng/mL at the start of thethird consecutive treatment cycle (e.g., prior to dosing on day 15 orCycle 3, Day 1), and the steady state blood plasma concentration ofCompound 1 in the patient being maintained at or above about 2,000 ng/mLand below about 7,500 ng/mL (preferably, below about 7,200 ng/mL)throughout the course of treatment. In some embodiments, the steadystate blood plasma concentration of Compound 1 in the patient ismaintained at or above about 1652 ng/mL and below about 7,840 ng/mLthroughout the course of treatment.

Maintaining Therapeutically Effective Blood Plasma Concentrations ofCompound 1 Throughout a Course of Treatment

The invention is based in part on the discovery that oral administrationof Compound 1 (in the pharmaceutically acceptable oral dosage formresulting from the preparation method of Example 1) to humans canprovide steady state (trough) blood concentrations above atherapeutically effective amount (e.g., above the IC₉₀ concentration forR132H and/or R132C mIDH-1, and/or concentrations of greater than about2,000 ng/mL) throughout a course of treatment of at least up to 30 weeksand beyond starting from the initial administration of Compound 1.Compound 1 can be administered to patients having elevated blood 2-HGlevels (i.e., above about 180 ng/mL), leading to a reduction in 2-HGlevels in the blood within 15 consecutive days starting with the firstday of the administration of Compound 1, followed by maintaining thelevels of 2-HG to below 200 ng/mL throughout the ensuing course oftreatment. Alternatively, Compound 1 can be administered to patientshaving elevated blood 2-HG levels (i.e., above about 180 ng/mL), leadingto a reduction in 2-HG levels in the blood within 15 consecutive daysstarting with the first day of the administration of Compound 1,followed by maintaining the levels of 2-HG to below 180 ng/mL throughoutthe ensuing course of treatment. Compound 1 was administered in an oraldosage form (obtainable from the method in Example 1) twice per day (150mg BID). After the initial 15 days of treatment with 150 mg BID of thisoral dosage form of Compound 1, the steady state (trough) bloodconcentration of Compound 1 (pre-dose) was maintained above about 2,000ng/mL and well below the predicted threshold for QTc risk (e.g., belowabout 7,200 ng/mL) throughout a course of treatment (e.g., at least upto about 30 weeks or longer, including 12-30 weeks or up to 6 months orlonger, from initial administration of Compound 1). Alternatively, afterthe initial 15 days of treatment with 150 mg BID of this oral dosageform of Compound 1, the steady state (trough) blood concentration ofCompound 1 (pre-dose) was maintained above about 1652 ng/mL and wellbelow the predicted threshold for QTc risk (e.g., below about 7840ng/mL) throughout a course of treatment (e.g., at least up to about 30weeks or longer, including 12-30 weeks or up to 6 months or longer, frominitial administration of Compound 1). In some embodiments of themethods of treatment disclosed herein, the steady state bloodconcentration of Compound 1 after day 15 is maintained at greater thanabout 10-times the measured blood concentrations of 2-HG in the patient(e.g., at or below about 200 ng/mL). A R132X mIDH-1 Selective InhibitorTherapy provides administering to a patient in need thereof a total doseof 150 mg BID of a pharmaceutically acceptable form of Compound 1provided in Example 1 in an oral dosage form, on consecutive daysthroughout a Course of Treatment. Compound 1 is preferably the onlyinhibitor of mutant IDH-1 (mIDH-1) having one or more R132X mIDH-1Mutation(s) administered to the patient throughout the Course ofTreatment of the R132X mIDH-1 Selective Inhibitor Therapy. Unlessotherwise indicated, the mIDH-1 selective inhibitor (e.g. Compound 1)can be administered as a single agent as the R132X mIDH-1 SelectiveInhibitor Therapy, or in combination with other therapeutic agents thatare not mIDH-1 inhibitors as a combination for the R132X mIDH-1Selective Inhibitor Therapy.

Compound 1 is a potent and selective small molecule inhibitor of certainmutated forms of the isocitrate dehydrogenase 1 (IDH-1) enzyme. Compound1 selectively inhibits mutant IDH-1 enzymes compared to the wild typeIDH-1 enzyme, targeting the mutant IDH-1 variants defined herein asR132X mIDH-1 Mutation(s). Example 2 provides in vitro data demonstratinginhibition of various R132X mutations of mIDH-1 enzyme. For example,Compound 1 targets the mutant IDH-1 variants R132H, R132C, R132L, R132G,and R132S using assays described in Example 2 with IC₅₀ concentrationsthat are approximately at least 180-fold lower than the wild-type IDH-1enzyme in vitro. In addition, Compound 1 targets the R132H and R132Cmutations of IDH-1 at IC₅₀ concentrations demonstrating selectivity overwild-type IDH-1 enzyme in vitro (based on IC₅₀ measurements asdetermined in Example 2 based on average +/−SEM, nM). Accordingly,preferred R132X mutations include R132H and R132C, as well as R132L,R132G, and R132S (or other R132X mutations having therapeuticallyrelevant 2-HG IC₅₀ values obtained using the in vitro assay of Example2). In addition, Compound 1 selectively inhibits mutant IDH-1 comparedto mutant IDH-2 forms. The selectivity of Compound 1 against other IDHisozymes was tested using diaphorase coupled assays employing eitherwild-type IDH-1 or one of 2 alternative mutated forms of IDH-2 (R140Qand R172K). Compound 1 had very weak activity against either wild typeIDH-1 or R172K IDH-2 mutation (with enzymatic IC₅₀ measurements obtainedaccording to Example 2 of about 20-25 micromolar, compared with IC₅₀values of less than about 150 nanomolar obtained for the R132X m-IDH-1Mutations). In addition, Compound 1 did not show any inhibition of R140QIDH-2 up to a concentration of 100 micromolar. These selectivity datademonstrate that Compound 1 is a potent and selective inhibitor ofenzymes harboring R132X mIDH-1 Mutation(s).

The R132X mIDH-1 Selective Inhibitor Therapy (single agent orcombination) can be administered to adult patients with an IDH-1mutation as detected by a medically appropriate (e.g., FDA-approved)test for mIDH-1 mutation(s). Preferably, the test is a diagnostic thatidentifies an R132X mIDH-1 Mutation(s) in the patient prior to theadministration of Compound 1. Preferably, the patient is identified asharboring one or more R132X mIDH-1 Mutation(s) based on Next GenerationSequencing (NGS) detection on a tissue sample obtained from the patientprior to the administration of Compound 1 and/or administration of anyR132X mIDH-1 Selective Inhibitor Therapy.

A patient in need of R132X mIDH-1 Selective Inhibitor Therapy can havean elevated level of 2-HG measured in the patient (e.g., in the bloodplasma of the patient) prior to initiating any R132X mIDH-1 SelectiveInhibitor Therapy. Preferably, the level of 2-HG measured in the bloodof the patient declines during the first 2 weeks of the Course ofTreatment of a R132X mIDH-1 Selective Inhibitor Therapy. For example, apatient may have a measured blood concentration level of 2-HG that isgreater than about 200 ng/mL of 2-HG prior to the administration ofCompound 1 pursuant to administration of a R132X mIDH-1 SelectiveInhibitor Therapy to the patient in need thereof, and a measured bloodconcentration level of 2-HG of less than about 200 ng/mL during a Courseof Treatment with a R132X mIDH-1 Selective Inhibitor Therapy. Forexample, in the human clinical trial disclosed in Example 3, all IDH-1m+patients had elevated 2-HG, which was reduced upon treatment withCompound 1 by day 15 of the Course of Treatment, with about 30%demonstrating a response to the administration of Compound 1 at somepoint during the Course of Treatment. In this patient population, thenormal 2-HG measured in patient blood at CRL was about 70±17 ng/mL; theobserved highest was about 91 ng/mL and the lowest was about 43 ng/mL.Preferably, the R132X mIDH-1 Selective Inhibitor Therapy consists of theadministration of Compound 1 (i.e., Compound 1 is the only mIDH-1inhibitor administered to the patient throughout the Course ofTreatment).

Compound 1 is administered over a therapeutically effective Course ofTreatment, which is preferably long enough to provide and sustain anintended therapeutic effect. For example, the Course of Treatment can belong enough to therapeutically reduce elevated 2-HG levels in a patient(e.g., reduce 2-HG levels measured in patient blood plasma to belowabout 200 ng/mL), with continued administration of Compound 1 to thepatient in a manner that provides therapeutically effective steady stateblood plasma concentration levels of Compound 1 (e.g., trough bloodplasma concentrations greater than the IC₉₀ concentration value for 2HGproduction measured for a R132X IDH-1 mutation identified in cellsobtained from the patient). When treating patients with elevated 2-HGlevels measured in the patient's blood prior to administration ofCompound 1, the Course of Treatment can be at least a number ofconsecutive days starting from the initial administration of Compound 1to the patient with elevated 2-HG levels, and continuing with dailyadministration of Compound 1 (e.g., 150 mg BID) for at least a number ofdays effective to reduce the 2-HG levels measured in the blood of thepatient to less than about 200 ng/mL (preferably less than 180 ng/mL)and/or a level considered medically appropriate for the patient (e.g.,to a range determined to be medically normal for that patient in thetreatment paradigm). Preferably, the Course of Treatment is at least 15consecutive days starting with the day of the initial administration ofCompound 1 to the patient and comprises 150 mg of the solid oral dosageform of Compound 1 (e.g., Example 1) administered to the patient twiceper day (e.g., every 12 hours) every day for at least 15 days. TheCourse of Treatment can be one or more 28-day treatment cycles of dailyBID administration of 150 mg of Compound 1 in the solid oral dosage formobtained from Example 1. The Course of Treatment can continue throughouta medically appropriate number of days for a patient. For example, theCourse of Treatment can last for any medically appropriate number ofconsecutive 28-day treatment cycles, including a Course of Treatmentlasting for 1, 2, 3, 4, 5, 6 consecutive treatment cycles of 28-dayseach, and/or a Course of Treatment of 20 weeks, and/or a Course ofTreatment of 6 months or more. In some methods, the Course of Treatmentis at least 6 months, or between at least 15 consecutive days and 6months of consecutive days of treatment comprising administration of 150mg BID of Compound 1 in a pharmaceutical form obtained from Example 1.

The R132X mIDH-1 Selective Inhibitor Therapy (e.g., oral administrationof 150 mg BID of Compound 1 throughout a Course of Treatment) providedunexpectedly durable blood concentration levels throughout the Course ofTreatment exceeding 6 months. The level of steady state bloodconcentration during the Course of Treatment was durable, meaning thatthe steady state blood concentration of Compound 1 did not decrease bymore than 10% throughout a Course of Treatment with continuedadministration of Compound 1 at a dose of 150 mg BID each day, whileremaining within a therapeutic concentration window defined by a minimumconcentration above the IC₉₀ determined in vitro for the 2-HG productionof a R132X mIDH-1 mutation harbored by the patient (e.g., above about2,000 ng/mL or above about 1652 ng/mL), and a maximum concentrationvalue below the concentration associated with medically unacceptablerisk of QTc prolongation (e.g., about 7,200 ng/mL or about 7840 ng/m L).

In addition, the R132X mIDH-1 Selective Inhibitor Therapy can reduce2-HG levels in the blood of the patient, although this reduction did notcorrelate with disease response in the patients during the Course ofTreatment. As shown in FIGS. 9A-9D, the R132X mIDH-1 Selective InhibitorTherapy provided a sustained reduction in 2HG levels, meaning thatelevated levels of the blood concentration of 2-HG in patient bloodplasma were reduced relative to pre-dose levels during the initialportion of a Course of Treatment (e.g., within the first 15 days) andthen sustained at or below about the pre-dose level throughout the restof the Course of Treatment (e.g., at or below about 200 ng/mL for 6consecutive 28-day treatment cycles or at or below about 180 ng/mL for 6consecutive 28-day treatment cycles).

FIGS. 8A-8E is a graph showing the steady state (trough) bloodconcentration measured in patients after administration of 150 mg ofCompound 1 BID, as described in the human clinical trial of Example 10.FIGS. 8A-8E are different graphs each showing the steady state bloodconcentration of Compound 1 measured in patient blood for a collectionof patients in the human clinical trial described in Example 10. FIGS.8A-8E are each graphs showing the measured steady state bloodconcentrations measured in each of individual patients included in thepopulation, at the indicated time points, in the human clinical trial ofExample 10. Notably, the steady state blood concentration of Compound 1remained above a therapeutic minimum value (e.g., the IC₉₀ value for2-HG production by at least one of the R132X mutation(s) identified inpatient's tissue prior to administration of Compound 1) throughout theCourse of Treatment (e.g., 30 weeks). In particular, the administrationof Compound 1 in the oral dosage form described in Example 1 at 150 mgBID resulted in a sustained therapeutically effective trough bloodplasma concentration above 2,000 ng/mL throughout a 30-week Course ofTreatment as shown in FIGS. 8A-8E (e.g., including after cycle 3 of a28-day treatment cycle, after week 20 and continuing to week 30). Thetrough concentration measurements of Compound 1 in patient blood did notdecline below a therapeutically effective level at each point measuredduring the Course of Treatment. Referring to FIGS. 9A-9D, theadministration of Compound 1 in the oral dosage form from Example 1 at150 mg BID in the human clinical trial of Example 10 resulted in asustained 2-HG level under 200 ng/mL in plasma throughout the Course ofTreatment ranging from cycle 2, day 1 through cycle 8, day 1 (each cyclerepresents 28 consecutive days of oral administration of 150 mg BID ofthe solid form of Compound 1 obtainable from the production method ofExample 1).

The invention is based in part on the discovery that a R132X mIDH-1Selective Inhibitor Therapy where Compound 1 is the only mIDH-1inhibitor administered (administration of Compound 1 at 150 mg BID)unexpectedly resulted in a steady state blood concentration that wasdurable (e.g., blood plasma steady state concentration of Compound 1remains within about 20% (or does not decrease by more than about 20%),and preferably remains within 10% (or does not decrease by more than10%) of the concentration measured the day after the initial 28-daycycle in a Course of Treatment). In addition, the administration ofCompound 1 as described in Example 10 reduced elevated 2-HGconcentrations in the blood of the patients within about 15 days andthen sustained patient blood concentrations of 2-HG at less than about200 ng/mL after about 15 days of a Course of Treatment.

The R132X mIDH-1 Selective Inhibitor Therapy can provide a sustainedratio of greater than about 10 (preferably greater than about 20) ofblood plasma concentration of Compound 1 (e.g., trough concentrationsmeasured pre-dose at concentrations of about 2,000 ng/mL or greater) to2-HG blood level (e.g, concentrations of plasma concentrations of about200 ng/mL or lower, including concentrations of about 100 ng/mL) aftercycle 3 day 1 (i.e., after BID doses administered over the initial 15consecutive days of treatment) of a 28-day treatment cycle. A plasmahalf-life of about 60 hours was estimated for Compound 1, with steadystate achieved by week 2 of the course of treatment. The steady stateblood concentrations of Compound 1 measured in the patients was abovethe IC₉₀ value for 2-HG inhibition in R132X mIDH-1 cells (described inthe Examples). As shown in FIGS. 8A-8E, the plasma exposures (steadystate blood plasma concentration) of Compound 1 were durable (i.e.,sustained) throughout the 30-week treatment duration. As shown in FIGS.9A-9D, the plasma 2-HG concentrations were reduced to the normal rangewithin 1 cycle (C2D1) and maintained throughout the treatment duration.No dose limiting toxicities of Compound 1 were observed during doseescalation studies, and the maximum tolerated dose (MTD) of Compound 1was not reached. Preferably, the R132X mIDH-1 Selective InhibitorTherapy can be administered to a patient that has not received any otherR132X mIDH-1 inhibitor compound.

As described in Example 10, FIG. 11A shows the ratio of the steady stateblood concentration of Compound 1, normalized to 1.0 using theconcentration measured on day 15 of a Course of Treatment, for a singlepatient who received 150 mg BID of the solid form of Compound 1obtainable from Example 1 throughout a Course of Treatment of over 300days (i.e., greater than 6 months). The steady state blood exposure(concentration) of Compound 1 varied from about 90-133% of theconcentration of Compound 1 measured in the patient at cycle 1, day 15,throughout the subsequent remainder of this Course of Treatment.

Optionally, a hypomethylating agent and/or a nucleic acid synthesisinhibitor can also be administered to the patient during the Course ofTreatment. Suitable agents that can also be administered during theCourse of Treatment include azacitabine and/or decitabine.

In some embodiments, a combination therapy of Compound 1 and azacitidinecan be administered for the treatment of patients with certain forms ofcancer (e.g., glioma) harboring IDH-1 mutations. For example, patientscan be administered Compound 1 daily (BID) in continuous 28-day cycles,in combination with azacitidine (administered at the dose of 75 mg/m²for 7 days IV/SC per every 28-day cycle). Azacitidine (also, azacytidineor AZA herein) is believed to exert its antineoplastic effects bycausing hypomethylation of DNA and direct cytotoxicity on abnormalhematopoietic cells in the bone marrow. Azacitidine can be administeredduring a Course of Treatment at a subcutaneous dose of 75 mg/m² dailyfor 7 days every 4 weeks. The azacitidine dose can be increased to 100mg/m² if no beneficial effect was seen after 2 treatment cycles. Thedose of azacitidine can be decreased and/or delayed based on hematologicresponse or evidence of renal toxicity. Azacitidine is indicated fortreatment of patients with the following myelodysplastic syndromesubtypes: refractory anemia or refractory anemia with ringedsideroblasts (if accompanied by neutropenia or thrombocytopenia orrequiring transfusions), refractory anemia with excess blasts,refractory anemia with excess blasts in transformation, and chronicmyelomonocytic leukemia. In some embodiments, a method of treatmentcomprises (a) the (e.g., oral) administration of a total of 150 mg ofCompound 1 BID to a patient throughout a Course of Treatment; and (b)administering a therapeutically effective amount of azacitidine to thepatient (e.g, administering azacitidine at a dose of 75 mg/m² daily for7 days every 4 weeks, wherein the azacitidine dose can be increased to100 mg/m² if no beneficial effect was seen after 2 treatment cycles andthe dose of azacitidine can be decreased and/or delayed based onhematologic response or evidence of renal toxicity).

Decacitabine (5-aza-2′-deoxycytidine) is a nucleoside metabolicinhibitor indicated for treatment of patients with myelodysplasticsyndromes (MDS) including previously treated and untreated, de novo andsecondary MDS of all French-American-British subtypes (refractoryanemia, refractory anemia with ringed sideroblasts, refractory anemiawith excess blasts, refractory anemia with excess blasts intransformation, and chronic myelomonocytic leukemia) and intermediate-1,intermediate-2, and high-risk International Prognostic Scoring Systemgroups. In some embodiments, a method of treatment comprises the (e.g.,oral) administration of a total of 150 mg of Compound 1 BID to a patientthroughout a Course of Treatment; and (b) administering atherapeutically effective amount of decacitabine to the patient (e.g,administering decacitabine at a dose of 15 mg/m² by continuousintravenous infusion over 3 hours repeated every 8 hours for 3 days andrepeating this cycle every 6 weeks; or administering the decacitabine ata dose of 20 mg/m² by continuous intravenous infusion over 1 hourrepeated daily for 5 days. Repeat cycle every 4 weeks).

Glioma

The present disclosure provides methods for treating solid tumors in theCNS, including a brain cancer tumor, harboring a R132 IDH-1 mutation.For example, patients diagnosed with brain cancer harboring a mutantIDH-1 cancer cell can be treated with a therapeutically effective amountof Compound 1 in combination with azacitidine.

Compound 1 is a small molecule inhibitor of mutated forms of isocitratedehydrogenase 1 (IDH-1) enzyme, and is useful for the treatment of adultpatients diagnosed with cancer having an IDH-1 mutation as detected byan FDA-approved test. Compound 1 can be administered to patients in needthereof in a therapeutically effective amount (e.g., 150 mg orally twicedaily until disease progression or unacceptable toxicity). Patients forthe treatment of cancer with Compound 1 can be selected based on thepresence of IDH-1 mutations in the blood or bone marrow. In oneembodiment, the recommended starting dose of Compound 1 is 150 mg takenorally twice daily with or without food until disease progression orunacceptable toxicity. For patients without disease progression orunacceptable toxicity, the patient can receive the therapeuticallyeffective amount of Compound 1 for a minimum of 6 months to allow timefor clinical response.

The invention is based in part on preclinical studies showing thatCompound 1 can cross the blood brain barrier (BBB) in mouse models. Oraladministration of Compound 1 showed high systemic bioavailabilty inmultiple preclinical species. Permeability was excellent, with littleevidence of efflux, and significant brain penetration was observed inmice (98% brain binding in murine animal model).

Preferably, patients diagnosed with glioma harboring a R132 IDH-1mutation can be treated with a therapeutically effective combination ofa pharmaceutical composition of Compound 1 (e.g., an oral dosage formproviding 150 mg of Compound 1 administered twice per day on consecutivedays for a course of treatment comprising multiple treatment cyclestotaling at least 6 months) and azacitidine. The azacitidine can besubcutaneously or intravenously administered to the patient in anazacitidine treatment cycle consisting of the administration of a totaldose of 75 mg/m² each day for 7 consecutive days beginning at the startof each treatment cycle, followed by 21 consecutive days withoutadministration of the azacitidine to the patient. A 48-hourdose-interruption of azacitidine is allowed for weekends or holidays. Ifno response is seen after 2 treatment cycles, azacitidine can beadministered at a total dose of 100 mg/m² each day.

Compound 1 is preferably administered on consecutive days throughout aCourse of Treatment. As used herein, the term “Course of Treatment”refers to the time period in which a patient is being administeredCompound 1 (e.g., as a single agent, or in combination with anothertherapeutic agent), including any administration holidays or recoveryperiods. A course of treatment can include a single treatment cycle ormultiple treatment cycles. For example, a Course of Treatment cancomprise one or more 28-day treatment cycles. Additionally, a course oftreatment can include a partial treatment cycle. The Course of Treatmentcan include the total time period during which a patient is on atreatment protocol for a therapy comprising the administration of amIDH-1 inhibitor compound. Preferably, the Course of Treatment is atleast about 15 consecutive days, at least about a 28-consecutive daytreatment cycle, or at least about four, five, six or more consecutive28-day treatment cycles and more preferably, at least about 4 months orlonger (e.g., 6 months or longer). Preferably, Compound 1 isadministered twice per day (e.g., about every 12 hours) every daythroughout a Course of Treatment.

In some embodiments, patients can be treated with Compound 1 incombination with a hypomethylating agent such as azacitidine ordecitabine. The recommended starting dose for azacitidine in the firsttreatment cycle, for all patients regardless of baseline hematologylaboratory values, is 75 mg/m² of body surface area, injectedsubcutaneously, daily for 7 days, followed by a rest period of 21 days(28-day treatment cycle). It is recommended that patients be treated fora minimum of 6 cycles. Treatment should be continued as long as thepatient continues to benefit or until disease progression. In somemethods, azacitidine is administered to the patient in need thereof at adose of 75 mg/m², SC, d1-7, q4 wk throughout a course of treatment,while receiving Compound 1 at a dose of 150 mg BID. In other methods,decitabine is administered to the patient in need thereof at a dose of20 mg/m², IV, d1-5, q4 wk, while receiving Compound 1 at a dose of 150mg BID.

In one embodiment, patients diagnosed with glioma harboring a mIDH1 canbe treated with a mIDH1 Inhibitor Therapy consisting of Compound 1 andazacitidine. Treatment with the hypomethylating agent azacitidine cancause tumor growth inhibition in a patient-derived IDH1-mutated gliomamodel by reducing DNA methylation and inducing glial differentiation.IDH1 R132H mutations represent more than 90% of the IDH mutationspresent in low grade glioma and secondary GBM patients. The IDH1mutations R132C and R132S are also reported in glioma patients. At leastin mIDH1 harboring cancer cells, wild type and mutant IDH1 form aheterodimeric complex that can produce very high 2-HG levels (up to 3-35mM in glioma cells). For example, patients bearing IDH1 mutations haveelevated levels of 2-HG, which in some cases reach tumorconcentrations >10 mM (glioma).

In another embodiment, patients diagnosed with chondrosarcoma harboringa mutant IDH1 cancer cell can be treated with a therapeuticallyeffective amount of Compound 1 alone or in combination with azacitidine.In some embodiments, a combination therapy comprising Compound 1 andazacitidine can be administered for the treatment of patients withchondrosarcoma harboring IDH1 mutations. For example, patients can beadministered Compound 1 daily (BID) in continuous 28-day cycles, incombination with azacitidine (administered at the daily dose of 75 mg/m²for 7 days IV/SC per every 28-day cycle).

Preferably, patients treated with a combination of Compound 1 andazacitidine receive a therapeutically effective amount of a mIDH1Inhibitor Therapy selected from a dose level indicated in Table 6 below.

TABLE 6 Preferred Dose Levels for mIDH1 Inhibitor Therapy withAzacitidine Dose Level Compound 1 Azacitidine 1 (Starting Dose) 150 mgBID 75 mg/m²/day × 7 continuously for 28 days every 28 days consecutivedays −1 (Hematologic 150 mg BID 37 mg/m²/day × 7 Dose-Limitingcontinuously for 28 days every 28 days Toxicity (DLT)) consecutive days−1 (non-Hematologic 150 mg BID 75 mg/m²/day × 7 DLT) continuously for 28days every 28 days consecutive days

Patients diagnosed with hepatobiliary carcinoma (HBC) harboring a mutantIDH1 cancer cell can be treated with a therapeutically effective amountof Compound 1 alone or in combination with a PD-1 inhibitor (e.g.,Pembrolizumab (Keytruda) or Nivolumab (Opdivo)). In some embodiments, acombination therapy of Compound 1 and the PD-1 inhibitor can beadministered for the treatment of patients with a HBC cancer harboringIDH1 mutations. For example, patients can be administered compound 1daily (BID) in continuous 28-day cycles, in combination withPembrolizumab (e.g., administered at the dose of 200 mg every 3 weeks).For example, patients can be administered compound 1 daily (BID) incontinuous 28-day cycles, in combination with Nivolumab (e.g.,administered at the dose of 240 mg every 2 weeks or 480 mg every 4weeks). Preferably, patients treated with a combination comprisingCompound 1 and Nivolumab receive a therapeutically effective amount of amIDH1 Inhibitor Therapy selected from a dose level indicated in Table 7below.

TABLE 7 Preferred Dose Levels for mIDH1 Inhibitor Therapy with NivolumabDose Level Compound 1 Nivolumab 1 (Starting 150 mg BID continuously 240mg intravenous Dose) for 28 consecutive days every 2 weeks −1 (any DLT)150 mg once daily 240 mg intravenous continuously for 28 every 2 weeksconsecutive days

Patients diagnosed with IHCC harboring a mutant IDH1 cancer cell can betreated with a therapeutically effective amount of Compound 1 alone orin combination with a chemotherapy (e.g., gemcitabine and cisplatin).Preferably, patients treated with a combination of Compound 1 andgemcitabine and cisplatin chemotherapy receive a therapeuticallyeffective amount of a mIDH1 Inhibitor Therapy selected from a dose levelindicated in Table 8 below.

TABLE 8 Preferred Dose Levels for mIDH1 Inhibitor Therapy withGemcitabine/Cisplatin Dose Level Compound 1 Gemcitabine/Cisplatin 1(Starting 150 mg BID Cisplatin 25 mg/m² Dose) continuously for 28followed by gemcitabine and Day 8 consecutive days 1,000 mg/m² on Day 1−1 (any DLT) 150 mg once daily Cisplatin 25 mg/m₂ followed bycontinuously for 28 gemcitabine 1,000 mg/m² consecutive days on Day 1and Day 8

It will be appreciated that use of headers in the present disclosure areprovided for the convenience of the reader. The presence and/orplacement of a header is not intended to limit the scope of the subjectmatter described herein.

The present disclosure contemplates, among other things, the followingnumbered embodiments:

-   1. A method of treating AML or MDS in a patient harboring isocitrate    dehydrogenase 1 mutations (mIDH-1), the method comprising    administering to a patient in need thereof a therapeutically    effective amount of a R132X mIDH-1 Selective Inhibitor Therapy    consisting of Compound 1 in an oral dosage form for a course of    treatment starting with the initial administration of Compound 1 and    continuing on consecutive days for at least 15 days, wherein the    administration of the therapeutically effective amount of Compound 1    results in the patient having a durable therapeutically effective    trough blood plasma concentration of Compound 1 in the patient    throughout the course of treatment.-   2. A method of treating AML or MDS in a patient harboring isocitrate    dehydrogenase 1 mutations (mIDH-1), the method comprising    administering to a patient in need thereof a therapeutically    effective amount of Compound 1 for at least three consecutive    treatment cycles of 28 consecutive days each, wherein the    administration of the therapeutically effective amount of Compound 1    results in the patient having both:    -   a. a durable sustained therapeutically effective trough blood        plasma concentration of Compound 1 in the patient throughout the        course of treatment, and    -   b. a reduced level of 2-HG in the patient's plasma after the        first two consecutive treatment cycles.-   3. The method of any one of embodiments 1 or 2, wherein    -   a. the steady state blood plasma concentration of Compound 1 in        the patient is maintained at or above about 2,000 ng/mL        throughout the course of treatment, and    -   b. the level of 2-HG in the patient plasma is maintained at or        below about 200 ng/mL after the start of the third consecutive        treatment cycle of the course of treatment on day 15 after the        initial dose of Compound 1.-   4. The method of any one of embodiments 1-3, wherein the Compound 1    is administered in a dose of 150 mg/day twice per day throughout the    course of treatment.-   5. The method of any one of embodiments 1-4, wherein the Compound 1    is administered with food to improve bioavailability of Compound 1.-   6. The method of any one of embodiments 1-5, wherein the course of    treatment is 12-30 weeks.-   7. The method of any one of embodiments 1-6, wherein the method    further comprises the administration of azacitidine to the patient    throughout the course of treatment.-   8. The method of any one of embodiments 1-6, wherein the patient is    receiving or previously received treatment with azacitidine.-   9. The method of embodiment 7 or 8, wherein the azacitidine is    subcutaneously or intravenously administered to the patient in an    azacitidine treatment cycle consisting of the administration of a    total dose of 75 mg/m² each day for 7 consecutive days beginning at    the start of each treatment cycle, followed 21 consecutive days    without administration of the azacitidine to the patient.-   10. The method of any one of embodiments 1-6, wherein the method    further comprises the administration of cytarabine to the patient    throughout the course of treatment.-   11. The method of any one of embodiments 1-6, wherein the patient is    receiving or previously received treatment with cytarabine.-   12. The method of embodiment 10 or 11, wherein the cytarabine is    subcutaneously or intravenously administered to the patient in a    cytarabine treatment cycle consisting of the administration of a    total dose of 20 mg/day each day for 7 consecutive days beginning at    the start of each treatment cycle, followed 10 consecutive days    without administration of the cytarabine to the patient.-   13. The method of any one of embodiment 1-12, wherein the patient    has been identified as having a R132X mutation in mIDH-1 using a    diagnostic method comprising a next generation sequencing (NGS)    analysis of a bone marrow or other tissue sample from the patient    obtained prior to the administration of Compound 1 to the patient.-   14. The method of any one of embodiments 1-13, wherein the subject    has relapsed or refractory (R/R) AML.-   15. The method of any one of embodiments 1-13, wherein the subject    has AML or MDS with residual R132X mIDH-1 in morphologic complete    remission or complete remission with incomplete blood count recovery    (CR/CRi) after cytotoxic-containing induction therapy with residual    IDH-R132X mutation.-   16. The method of any one of embodiments 1-13, wherein the subject    has relapsed or refractory AML or MDS previously treated with an    IDH1 inhibitor.-   17. The method of any one of embodiments 1-13, wherein the subject    has relapsed or refractory AML or MDS that are naïve to prior    hypomethylating therapy and IDH1 inhibitor therapy-   18. The method of any one of embodiments 1-13, wherein the subject    has relapsed or refractory AML or MDS that has inadequately    responded or has progressed immediately preceding hypomethylating    therapy.-   19. The method of any one of embodiments 1-13, wherein the subject    is a subject with relapsed or refractory AML or MDS that have been    previously treated with single-agent IDH1 inhibitor therapy as their    last therapy prior to study enrollment.-   20. The method of any one of embodiments 1-19, wherein the subject    has been diagnosed with AML with a R132X mIDH-1.-   21. The method of any one of embodiments 1-19, wherein the subject    has MDS with AML with a R132X mIDH-1.-   22. The method of any one of embodiments 1-21, wherein the subject    has been diagnosed with a R132X mIDH-1 selected from the group    consisting of R132H, R132C or both R132H and R132C.-   23. A method of treating AML or MDS in a patient harboring at least    one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method    comprising administering to the patient in need thereof a R132X    mIDH-1 Selective Inhibitor Therapy consisting of 150 mg BID of    Compound 1 in an pharmaceutically acceptable oral dosage form for a    course of treatment starting with the initial administration of    Compound 1 and continuing on consecutive days for at least 15 days.-   24. The method of embodiment 23, wherein the course of treatment    comprises at least 4 consecutive treatment cycles each consisting of    28 consecutive days of administering Compound 1 to the patient twice    per day.-   25. The method of embodiment 23, wherein the course of treatment    comprises 12-30 weeks of consecutive days of administering Compound    1 to the patient twice per day.-   26. A method of treating AML or MDS in a patient harboring at least    one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method    comprising administering to the patient 150 mg BID of Compound 1 in    a pharmaceutically acceptable oral dosage form for a course of    treatment starting with the initial administration of Compound 1 and    continuing on consecutive days for at least 15 days.-   27. A method of treating AML or MDS in a patient harboring at least    one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method    comprising administering to the patient 150 mg BID of Compound 1 in    a pharmaceutically acceptable oral dosage form for a course of    treatment starting with the initial administration of Compound 1 and    continuing on consecutive days for at least 12 weeks.-   28. A method of treating AML or MDS in a patient harboring at least    one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method    comprising administering to the patient 150 mg BID of Compound 1 in    a pharmaceutically acceptable oral dosage form for a course of    treatment starting with the initial administration of Compound 1 and    continuing on consecutive days for at least 30 weeks.-   29. A method of treating AML or MDS in a patient harboring one or    more R132X isocitrate dehydrogenase 1 mutations (mIDH-1), the method    comprising administering to a patient in need thereof a    therapeutically effective amount of a R132X mIDH-1 Selective    Inhibitor Therapy consisting of Compound 1 in an oral dosage form    for a course of treatment starting with the initial administration    of Compound 1 and continuing on consecutive days for at least 15    days, wherein the administration of the therapeutically effective    amount of Compound 1 results in the patient having both:    -   a. a durable trough blood plasma concentration of Compound 1 in        the patient measured at or above the IC90 concentration of        Compound 1 for 2-HG suppression of the R132X mIDH-1 throughout        the course of treatment, and    -   b. a level of 2-HG in the patient's plasma of less than about        200 ng/mL within two initial consecutive treatment cycles that        is maintained throughout the course of treatment.-   30. The method of embodiment 29, wherein course of treatment is at    least 3 consecutive 28-day treatment cycles.-   31. The method of embodiments 29 or 30, wherein the trough blood    plasma concentration of Compound 1 in the patient is measured    between 2,000 ng/mL-7,200 ng/mL.-   32. The method of any one of embodiments 29-31, wherein the level of    2-HG is maintained at about 180 ng/mL in the blood plasma of the    patient throughout the course of treatment after the first 15    consecutive days of administering Compound 1 to the patient.-   33. The method of any one of embodiments 1-32, wherein the    concentration of Compound 1 measured in the blood of the patient    does not decline more than 20% throughout the course of treatment    compared to the steady state blood concentration of Compound 1    measured prior to administration of Compound 1 at cycle 2, day 1    (day 29).-   34. The method of any one of embodiments 1-32, wherein the    concentration of Compound 1 measured in the blood of the patient    does not decline more than 10% throughout the course of treatment    compared to the steady state blood concentration of Compound 1    measured prior to administration of Compound 1 at cycle 2, day 1    (day 29).-   35. The method of any one of embodiments 33 and 34, wherein the    course of treatment is a total of 12 weeks.-   36. The method of any one of embodiments 33 and 34, wherein the    course of treatment is a total of 30 weeks.-   37. The method of any one of embodiments 22-36, wherein the subject    has been diagnosed with a R132X mIDH-1 selected from the group    consisting of R132H, R132C or both R132H and R132C.-   38. The method of any one of embodiments 1-37, wherein Compound 1 is    administered in the pharmaceutically acceptable dosage form    obtainable from Step 6 of Example 1.-   39. A method of treating AML in a patient harboring at least one    R132X isocitrate dehydrogenase 1 mutation (mIDH-1) selected from the    group consisting of: R132C, R132H, R132S, R132G, and R132L, wherein    the method comprises administering to the patient 150 mg BID of    Compound 1 in a pharmaceutically acceptable oral dosage form for a    course of treatment starting with the initial administration of    Compound 1 and continuing for at least 28 consecutive days.-   40. A method of treating AML in a patient harboring one or more    R132X isocitrate dehydrogenase 1 mutations (mIDH-1) selected from    the group consisting of: R132C, R132H, R132S, R132G, and R132L,    wherein the method comprises administering to a patient in need    thereof a therapeutically effective amount of a R132X mIDH-1    Selective Inhibitor Therapy consisting of Compound 1 in an oral    dosage form for a course of treatment starting with the initial    administration of Compound 1 and continuing for at least 15    consecutive days, wherein the administration of the therapeutically    effective amount of Compound 1 results in the patient having a    durable trough blood plasma concentration of Compound 1 in the    patient measured at or above the IC₉₀ concentration of Compound 1    for 2-HG suppression of the R132X mIDH-1 throughout the course of    treatment.-   41. The method of embodiment 40, wherein the administration of the    therapeutically effective amount of Compound 1 results in the    patient having a level of 2-HG in the patient's plasma of less than    about 200 ng/mL within two initial consecutive treatment cycles that    is maintained throughout the course of treatment.-   42. A method of treating AML in a patient harboring one or more    R132X isocitrate dehydrogenase 1 mutations (mIDH-1) selected from    the group consisting of: R132C, R132H, R132S, R132G, and R132L,    wherein the method comprises administering to a patient in need    thereof a therapeutically effective amount of a R132X mIDH-1    Selective Inhibitor Therapy consisting of Compound 1 in an oral    dosage form for a course of treatment starting with the initial    administration of Compound 1 and continuing for at least 15    consecutive days, wherein the administration of the therapeutically    effective amount of Compound 1 results in the patient having a level    of 2-HG in the patient's plasma of less than about 200 ng/mL within    two initial consecutive treatment cycles that is maintained    throughout the course of treatment.-   43. The method of embodiment 42, wherein the administration of the    therapeutically effective amount of Compound 1 results in the    patient having a durable trough blood plasma concentration of    Compound 1 in the patient measured at or above the IC₉₀    concentration of Compound 1 for 2-HG suppression of the R132X mIDH-1    throughout the course of treatment.-   44. A method of treating AML in a patient harboring one or more    R132X isocitrate dehydrogenase 1 mutations (mIDH-1) selected from    the group consisting of: R132C, R132H, R132S, R132G, and R132L, the    method comprising administering to a patient in need thereof a    therapeutically effective amount of a R132X mIDH-1 Selective    Inhibitor Therapy consisting of Compound 1 in an oral dosage form    for a course of treatment starting with the initial administration    of Compound 1 and continuing for at least 15 consecutive days,    wherein the administration of the therapeutically effective amount    of Compound 1 results in the patient having a durable trough blood    plasma concentration of Compound 1 in the patient measured at or    above 2,000 ng/mL throughout the course of treatment of at least 6    months.-   45. The method of embodiment 44, wherein the administration of the    therapeutically effective amount of Compound 1 results in the    patient having a level of 2-HG in the patient's plasma of less than    about 200 ng/mL within two initial consecutive 28-day treatment    cycles and is maintained throughout the course of treatment.-   44. A method of treating AML in a patient harboring one or more    R132X isocitrate dehydrogenase 1 mutations (mIDH-1) selected from    the group consisting of: R132C, R132H, R132S, R132G, and R132L,    wherein the method comprises administering to a patient in need    thereof:    -   a. a therapeutically effective amount of Compound 1 in an oral        dosage form for a Course of Treatment starting with the initial        administration of Compound 1 and continuing for at least 15        consecutive days for one or more consecutive R132X mIDH-1        Selective Inhibitor 28-day Treatment Cycles, wherein Compound 1        is the only R132X mIDH-1 selective inhibitor administered to the        patient during the Course of Treatment; and    -   b. azacitidine subcutaneously or intravenously administered to        the patient throughout the one or more R132X mIDH-1 Selective        Inhibitor 28-day Treatment Cycle(s), in an azacitidine treatment        cycle consisting of the administration of a total dose of 75        mg/m² each day for 7 consecutive days beginning at the start of        each treatment cycle, followed 21 consecutive days without        administration of the azacitidine to the patient;    -   wherein the administration of the therapeutically effective        amount of Compound 1 results in the patient having a durable        trough blood plasma concentration of Compound 1 in the patient        that is no less than about 90% of the concentration of Compound        1 measured after the first 15 days of the first R132X mIDH-1        Selective Inhibitor 28-day Treatment Cycle, throughout the        Course of Treatment of at least 6 months.-   45. A method of treating an adult patient diagnosed with a mutant    IDH-1 form of AML, comprising administering to the patient in need    thereof a combination therapy throughout a Course of Treatment    lasting at least 6 months, wherein the combination therapy comprises    a combination of:    -   a. 150 mg BID of Compound 1 each day throughout a Course of        Treatment of at least 6 months, wherein the Compound 1 is the        only mutant IDH-1 inhibitor administered to the patient during        the Course of Treatment;    -   b. azacitidine subcutaneously or intravenously administered to        the patient throughout the Course of Treatment at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.-   46. A method of treating an adult patient diagnosed with a mutant    IDH-1 form of AML, comprising administering to the patient in need    thereof 150 mg BID of Compound 1 each day throughout a Course of    Treatment of at least 6 months, in combination with azacitidine for    the treatment of the AML in the adult patient, wherein Compound 1 is    the only mutant IDH-1 inhibitor targeting R132C, R132H, R132S,    R132G, or R132L variants of IDH-1 that is administered to the    patient during the Course of Treatment.-   47. The method of embodiment 46, wherein the azacitidine is    administered subcutaneously or intravenously to the patient    throughout the Course of Treatment at a total dose of 75 mg/m² each    day for 7 consecutive days beginning at the start of each treatment    cycle, followed 21 consecutive days without administration of the    azacitidine to the patient.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient harboring one or more R132X    isocitrate dehydrogenase 1 mutations (mIDH-1), the method comprising    administering to the patient in need thereof a R132X mIDH-1    Selective Inhibitor Therapy consisting of the oral administration of    Compound 1 to the patient at a dose of 150 mg twice per day (BID)    for a Course of Treatment starting with the initial administration    of Compound 1 and continuing on consecutive days for at least 15    days.-   2. The method of embodiment 1, wherein Compound 1 is administered    with food to improve bioavailability of Compound 1.-   3. The method of any one of embodiments 1-2, wherein the Course of    Treatment is at least 12 weeks.-   4. The method of any one of claims 1-2, wherein the Course of    Treatment is at least 20 weeks.-   5. The method of any one of embodiments 1-2, wherein the Course of    Treatment is at least 30 weeks.-   6. The method of any one of embodiments 1-2, wherein the Course of    Treatment is at least 6 months.-   7. The method of any one of embodiments 1-6, wherein the subject has    a R132X mIDH-1 selected from the group consisting of R132H, R132C or    both R132H and R132C.-   8. The method of any one of embodiments 1-7, wherein Compound 1 is    administered in the solid form obtained from Example 1.-   9. A method of treatment of a disease in a patient harboring a    mIDH-1 mutation, the method comprising the steps of:-   a. selecting a patient for treatment based on the presence of an    IDH-1 mutation detected in cells obtained from the patient;-   b. administering Compound 1 to the selected patient from step (a) at    a starting dose of 150 mg taken orally twice daily until disease    progression or unacceptable toxicity.-   10. The method of embodiment 9, further comprising administering    Compound 1 to the selected patient for at least 6 months.-   11. The method of any one of embodiments 9-10, wherein Compound 1 is    administered to the patient as a single agent or in combination with    other therapeutic agents.-   12. The method of any one of embodiments 9-11, further comprising    assessing blood counts and blood chemistries of the selected patient    for leukocytosis and tumor lysis syndrome prior to the initiation of    Compound 1 in step (b).-   13. The method of any one of embodiments 9-12, further comprising    the step (c) of monitoring a selected patient at a minimum of every    2 weeks for at least the first 3 months during treatment of the    patient with compound 1 according to step (b).-   14. The method of any one of embodiments 9-13, wherein Compound 1 is    administered in the solid form obtained from Example 1 in a    pharmaceutically acceptable oral dosage form.-   15. The method of any one of embodiments 9-14 wherein the patient is    selected for treatment based on the detection of one or more R132X    IDH-1 mutations in cells obtained from the patient.-   16. The method of embodiment 15, wherein the R132X IDH-1 mutation is    detected using next generation sequencing of patient bone marrow    tissue.-   17. The method of any one of embodiments 15-16, wherein the R132X    mutation is selected from the group consisting of R132H and R132C.-   18. The method of any one of embodiments 16-17, wherein the R132X    IDH-1 mutation is detected prior to the administration of Compound    1.-   19. A method of treating a patient harboring one or more R132X    isocitrate dehydrogenase 1 mutations (m/DH-1), the method comprising    orally administering to the patient 150 mg of an oral pharmaceutical    dosage form of the compound    5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile    twice per day (BID) on consecutive days for at least 15 days.-   20. The method of embodiment 19, wherein the oral pharmaceutical    dosage form is the solid form of Compound 1 obtained by the process    of Example 1.-   21. The method of any one of embodiments 19-20, wherein the R132X    mutation is selected from the group consisting of R132H and R132C.-   22. The method of any one of embodiments 19-21, wherein the compound    is administered to the patient on consecutive days for at least 20    weeks.-   23. The method of any one of embodiments 19-22, wherein the compound    is administered to the patient on consecutive days for at least 30    weeks.-   24. The method of any one of embodiments 19-23, further comprising    detection of the R132X IDH-1 mutation in the patient using next    generation sequencing of patient bone marrow tissue prior to the    administration of the compound to the patient.-   25. A method of treatment comprising orally administering to a    patient harboring a R132X mIDH-1 arginine mutation a dose of 150 mg    BID of a pharmaceutically acceptable solid form of compound of    formula (I) as obtained from Example 1

-   26. The method of embodiment 25, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 20 weeks.-   27. The method of embodiment 25, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 30 weeks.-   28. The method of embodiment 25, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 6 months.-   29. The method of any one of embodiments 25-28, wherein Compound 1    is administered to the patient as a single agent or in combination    with other therapeutic agents.-   30. The method of any one of embodiments 25-29, wherein the R132X    mIDH-1 arginine mutation is selected from the group consisting of:    R132H, R132C, R132G, R132L, and R132S.-   31. The method of any one of embodiments 25-30, further comprising    detection of the R132X IDH-1 arginine mutation in the patient using    next generation sequencing of patient bone marrow tissue prior to    the administration of Compound 1 to the patient.-   32. The method of any one of embodiments 1-31, wherein the patient    has not received another mIDH-1 inhibitor compound.-   33. The method of any one of embodiments 1-8, wherein the patient    has greater than 200 ng/mL of 2-HG prior to initiating the R132X    mIDH-1 Selective Inhibitor Therapy.-   34. The method of any one of embodiments 9-18, wherein the patient    selected in step (a) has greater than 200 ng/mL of 2-HG prior to    administering Compound 1 in step (b).-   35. The method of any one of embodiments 19-24, wherein the patient    has greater than 200 ng/mL of 2-HG prior to administration of the    oral pharmaceutical dosage form of the compound to the patient.-   36. The method of any one of embodiments 25-31, wherein the patient    has greater than 200 ng/mL of 2-HG prior to administration of the    compound of formula (I) to the patient.-   37. A method of treatment comprising orally administering to a    patient having elevated blood levels of 2-HG greater than about 200    ng/mL and harboring at least one of a R132H, R132C, R132G, R132L,    and R132S IDH-1 mutation, a dose of 150 mg BID each day of a    pharmaceutically acceptable solid form of compound of formula (I) as    obtained from Example 1

-   -   on consecutive days for at least 15 days.

-   38. The method of embodiment 37, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 20 weeks.

-   39. The method of embodiment 37, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 30 weeks.

-   40. The method of embodiment 37, wherein the pharmaceutically    acceptable solid form of Compound 1 is administered to the patient    on consecutive days for at least 6 months.

-   41. A method of treating a patient diagnosed with a cancer harboring    one or more R132X isocitrate dehydrogenase 1 mutations (m/DH-1)    selected from the group consisting of R132C, R132H, R132L, R132G,    and R132S, the method comprising the oral administration of Compound    1 as the only inhibitor of mIDH-1 to the patient, at a total dose of    150 mg twice per day (BID) throughout a Course of Treatment starting    with the initial administration of Compound 1 and continuing on    consecutive days for at least 6 months.

-   42. A method of treating an adult patient diagnosed with a cancer    harboring one or more R132X isocitrate dehydrogenase 1 mutations    (m/DH-1) selected from the group consisting of R132C, R132H, R132L,    R132G, and R132S, the method comprising the oral administration of a    total dose of 150 mg twice per day (BID) of Compound 1 in the solid    form obtained from Example 1, on consecutive days throughout a    Course of Treatment starting with the initial administration of    Compound 1 and continuing on consecutive days for at least 6 months,    wherein Compound 1 is the only inhibitor of mIDH-1 administered to    the patient during a Course of Treatment.

-   43. The method of any one of embodiments 41-42, further comprising    the administration of an hypomethylating agent to the patient during    the Course of Treatment.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a glioma cancer    harboring a cancer cell with an IDH-1 R132 mutation, the method    comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 and a hypomethylating    agent over a course of treatment comprising multiple 28-day    treatment cycles.-   2. A method of treatment comprising the steps of:    -   a. selecting a patient diagnosed with a glioma cancer harboring        an IDH-1 mutation;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        for a treatment cycle of 28 consecutive days; and    -   c. administering azacitidine subcutaneously or intravenously to        the patient during the same treatment cycle as step (b), in a        total dose of 75 mg/m2 each day for the first 7 consecutive days        of the treatment cycle followed by 21 consecutive days without        the administration of azacitidine until the end of the treatment        cycle, with the exception of optionally permitting a 48-hour        dose interruption of azacitidine on a weekend of holiday during        the treatment cycle.-   3. The method of any one of embodiments 1-2, wherein the cancer does    not harbor a IDH-2 mutation.-   4. The method of any one of embodiments 1-3, wherein the patient has    been diagnosed as having a R132 mutation based on a patient    diagnostic.-   5. The method of embodiment 4, wherein the patient diagnostic    comprises detecting the R132 mutation in a biological sample    obtained from the patient.-   6. The method of embodiment 5, wherein the tissue sample is obtained    from the CNS of the patient.-   7. The method of any one of embodiments 4-6, wherein the R132    mutation is detected using next generation sequencing (NGS) without    the use of PCR.-   8. The method of any one of embodiments 1-7, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   9. The method of any one of embodiments 1-8, wherein Compound 1 is    administered to the patient once every 12 hours on consecutive days    throughout a course of treatment.-   10. The method of any one of embodiments 1-9, wherein Compound 1 is    administered to the patient throughout a course of treatment of at    least 6 months.-   11. The method of any one of embodiments 2-9, wherein Compound 1 is    administered to the patient throughout a course of treatment of at    least 6 consecutive treatment cycles.-   12. A method of treating a glioma cancer having an IDH-1 mutation in    an adult patient, the method comprising administering to the patient    in need thereof    -   a. a pharmaceutical composition comprising a total of 150 mg of        a pharmaceutically acceptable solid form of        5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile        obtained from Example 1, twice per day on consecutive days for a        28 day treatment cycle; and    -   b. administering azacitidine subcutaneously or intravenously to        the patient during the same treatment cycle as step (a), in a        total dose of 75 mg/m² each day for the first 7 consecutive days        of the treatment cycle followed by 21 consecutive days without        the administration of azacitidine until the end of the treatment        cycle, with the exception of optionally permitting a 48-hour        dose interruption of azacitidine on a weekend of holiday during        the treatment cycle.-   13. A method of treating a glioma cancer having an IDH-1 mutation in    an adult patient, the method comprising administering to the patient    in need thereof    -   a. a pharmaceutical composition comprising a total of 150 mg of        a Compound 1

-   -   in a pharmaceutically acceptable solid form (e.g. that        obtainable from Example 1), twice per day on consecutive days        for a 28 day treatment cycle; and    -   b. administering azacitidine subcutaneously or intravenously to        the patient during the same treatment cycle as step (a), in a        total dose of 75 mg/m² each day for the first 7 consecutive days        of the treatment cycle followed by 21 consecutive days without        the administration of azacitidine until the end of the treatment        cycle, with the exception of optionally permitting a 48-hour        dose interruption of azacitidine on a weekend of holiday during        the treatment cycle.

-   14. The method of any one of embodiments 1-13, wherein the patient    is diagnosed with glioblastoma multiforme prior to the    administration of Compound 1.

-   15. The method of any one of embodiments 1-14, wherein the patient    has elevated 2HG blood levels prior to the administration of    Compound 1.

-   16. The method of embodiment 15, wherein the level of 2HG measured    in the blood of the patient is greater than about 200 ng/mL prior to    the administration of Compound 1.

-   17. The method of any one of embodiments 1-16, wherein the level of    2HG measured in the blood of the patient is less than about 100    ng/mL measured on day 15 after the administration of Compound 1 for    the first 14 consecutive days of the first treatment cycle.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a cancer harboring    a cancer cell with an IDH-1 R132 mutation selected from the group    consisting of: R132L, R132G, and R132S, the method comprising    administering to the patient in need thereof a therapeutically    effective amount of Compound 1.-   2. The method of embodiment 1, wherein the cancer does not harbor a    IDH-2 mutation.-   3. The method of embodiment 1, wherein the cancer does not harbor a    IDH-2 mutation selected from the group consisting of: IDH-2 R172K    and IDH-2 R140Q.-   4. The method of embodiment 1, wherein the patient is diagnosed as    having a R132 mutation based on a patient diagnostic.-   5. The method of embodiment 4, wherein the patient diagnostic    comprises detecting the R132 mutation in a tissue sample obtained    from the patient.-   6. The method of embodiment 5, wherein the tissue sample is obtained    from the bone marrow of the patient.-   7. The method of any one of embodiments 4-6, wherein the R132    mutation is detected using next generation sequencing (NGS) without    the use of PCR.-   8. A method of treatment comprising the steps of:    -   a. selecting a patient for treatment based on the presence of        one or more IDH-1 mutations selected from the group consisting        of: R132L, R132G, and R132S;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        until disease progression or unacceptable toxicity.-   9. The method of embodiment 8, where the IDH-1 mutation is detected    in cancer cells obtained from the blood or bone marrow of the    patient.-   10. The method of embodiment 9, wherein the IDH-1 mutation is    detected prior to administering Compound 1 to the patient.-   11. The method of any one of embodiments 1-10, comprising the step    of detecting the IDH-1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   12. The method of any one of embodiments 1-11, wherein    administration of Compound 1 to the patient results in a decreased    2-hydroxyglutarate (2-HG) levels in the blood of the patient within    the first 15 consecutive days of treatment of the patient with    Compound 1.-   13. The method of any one of embodiments 1-12, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   14. The method of any one of embodiments 1-12, wherein the method    comprises administering 150 mg of Compound 1 to the patient twice    daily throughout a course of treatment.-   15. The method of embodiment 14, wherein the course of treatment is    at least 15 consecutive days.-   16. The method of any one of embodiments 1-15, wherein Compound 1 is    administered to the patient once every 12 hours on consecutive days    throughout a course of treatment.-   17. The method of any one of embodiments 1-16, wherein Compound 1 is    administered to the patient throughout a course of treatment of at    least 6 months.-   18. A method of inhibiting the production of 2-HG from a cell    harboring a IHD-1 mutation selected from the group consisting of:    R132L, R132G and R132S, the method comprising contacting the cell    with Compound 1 in an amount, under conditions, and for a time    sufficient to inhibit the production of 2-HG from the cell.-   19. A method of treating a patient diagnosed with a cancer harboring    a cancer cell with an IDH-1 R132 mutation, the method comprising    administering to the patient in need thereof a therapeutically    effective amount of Compound 1.-   20. The method of embodiment 19, wherein the patient is diagnosed    with a cancer harboring an IDH-1 R132 mutation in a cell obtained    from the patient, prior to the administration of Compound 1.-   21. A method of treating a patient diagnosed with a cancer harboring    a cancer cell with an IDH-1 R132 mutation, the method comprising    administering to the patient in need thereof a therapeutically    effective amount of Compound 1.-   22. The method of embodiment 21, wherein the patient is diagnosed    with a cancer harboring an IDH-1 R132 mutation in a cell obtained    from the patient, prior to the administration of Compound 1.-   23. A method of treating a patient diagnosed with a cancer, the    method comprising    -   a. diagnosing the patient as having a mutant IDH-1 mutation in a        cell obtained from the patient; and    -   b. administering a therapeutically effective amount of a        pharmaceutical composition comprising Compound 1 to the patient        in need of an inhibitor of the mutant IDH-1 enzyme that targets        the mutant IDH-1 variants R132C at no greater than about 5 times        the level of R132H; and    -   c. continuing to administer the pharmaceutical composition to        the patient throughout a course of treatment of at least 6        months.-   24. The method of embodiment 23, wherein the patient is in need of    an inhibitor of mIDH-1 variants selected from the group consisting    of R132L, R132G, and R132S;-   25. The method of any one of embodiments 23-24, wherein the relative    targeting of R132C and R132H variants of mIDH-1 is measured by the    ratio of IC₅₀ values obtained using the assay of Example 2.-   26. The method of any one of embodiments 23-25, wherein the patient    is diagnosed as having an IDH-1 mutation in a cell from the patient    using a next-generation sequencing (NGS)-based tumor genotyping    assay.-   27. The method of any one of embodiments 23-26, wherein the    pharmaceutical composition is administered to the patient twice per    day.-   28. The method of any one of embodiments 23-27, wherein the    pharmaceutical composition is administered to the patient in a dose    of 150 mg BID on consecutive days throughout the course of    treatment.-   29. The method of any one of embodiments 23-28, wherein Compound 1    in the pharmaceutical composition has the solid form obtained from    Example 1.-   30. A method of inhibiting the production of inhibiting the    production of 2-HG in a R132C mutated IDH-1 enzyme at no more than    about 5 times the inhibition of 2-HG production in a R132H mutated    IDH-1 enzyme, the method comprising contacting an IDH-1 enzyme not    having arginine at position 132 with a composition comprising    Compound 1 under conditions and for a time effective to inhibit 2-HG    production in either an IDH-1 R132C or an IDH-1 R132H mutation in    the mIDH-1 enzyme.-   31. A method of treating a cancer in an adult patient, the cancer    having a known mIDH-1 frequency of about 10-90%, the method    comprising administering to a patient diagnosed with an IDH-1    mutation comprising an IDH-1 mutation selected from the group    consisting of R132C, R132H, R132L, R132G, and R132S, the method    comprising administering to the patient in need thereof a    pharmaceutical composition comprising a total of 150 mg of a    pharmaceutically acceptable solid form of    5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile,    twice per day on consecutive days for a course of treatment    comprising 6 months.-   32. The method of embodiment 31, wherein the patient is diagnosed as    having an IDH-1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   33. The method of any one of embodiments 31-32, wherein the    pharmaceutical composition is administered to the patient every 12    hours.-   34. The method of any one of embodiments 31-33, wherein Compound 1    in the pharmaceutical composition has the solid form obtained from    Example 1.-   35. A method of treating a chrondrosarcoma cancer having an IDH-1    mutation in an adult patient, the method comprising administering to    the patient in need thereof a pharmaceutical composition comprising    a total of 150 mg of a pharmaceutically acceptable solid form of    5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile,    twice per day on consecutive days for a course of treatment    comprising 6 months.-   36. The method of embodiment 35, wherein the patient is diagnosed as    having an IDH-1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   37. The method of any one of embodiments 35-36, wherein the    pharmaceutical composition is administered to the patient every 12    hours.-   38. The method of any one of embodiments 35-37, wherein Compound 1    in the pharmaceutical composition has the solid form obtained from    Example 1.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a non-CNS solid    tumor harboring a cancer cell with an IDH1 R132 mutation, the method    comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 as a single agent.-   2. A method of treating a patient diagnosed with glioma harboring a    cancer cell with an IDH1 R132 mutation, the method comprising    administering to the patient in need thereof a therapeutically    effective amount Compound 1 in combination with a therapeutically    effective amount of azacitidine.-   3. A method of treating a patient diagnosed with a chondrosarcoma    cancer harboring a cancer cell with an IDH1 R132 mutation, the    method comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 in combination with a    therapeutically effective amount of azacitidine.-   4. A method of treating a patient diagnosed with a hepatobiliary    cancer harboring a cancer cell with an IDH1 R132 mutation, the    method comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 in combination with a    therapeutically effective amount of a PD-1 inhibitor.-   5. A method of treating a patient diagnosed with an intrahepatic    cholangiocarcinoma cancer harboring a cancer cell with an IDH1 R132    mutation, the method comprising administering to the patient in need    thereof a therapeutically effective amount of Compound 1 in    combination with a therapeutically effective amount of a gemcitabine    and cisplatin chemotherapy.-   6. The method of any one of embodiments 1-5, wherein Compound 1 is    administered at a dose of 150 mg taken orally twice daily.-   7. The method of any one of embodiments 1-6, wherein Compound 1 is    orally administered as the solid form obtained from Example 1.-   8. The method of any one of embodiments 1-7, further comprising the    steps of:    -   a. selecting a patient diagnosed with the cancer harboring an        IDH1 mutation;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        for a treatment cycle of 28 consecutive days.-   9. A method of treating a solid tumor or CNS cancer having an IDH1    mutation in an adult patient, the method comprising administering to    the patient in need thereof a pharmaceutical composition comprising    a total of 150 mg of a pharmaceutically acceptable solid form of    5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile    obtained from Example 1, twice per day on consecutive days for a 28    day treatment cycle.-   10. The method of any one of embodiments 1-9, wherein the cancer    does not harbor a IDH2 mutation.-   11. The method of any one of embodiments 1-10, wherein the cancer    does not harbor a IDH2 mutation selected from the group consisting    of: IDH2 R172K and IDH2 R140Q.-   12. The method of any one of embodiments 1-11, comprising the step    of detecting the IDH1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   13. The method of any one of embodiments 1-12, wherein    administration of Compound 1 to the patient results in a decreased    2-hydroxyglutarate (2-HG) levels in the blood of the patient after    the first 15 consecutive days of treatment of the patient with    Compound 1.-   14. The method of any one of embodiments 1-13, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   15. The method of any one of embodiments 1-14, wherein the method    comprises administering 150 mg of Compound 1 to the patient twice    daily throughout a course of treatment.-   16. The method of embodiment 15, wherein the course of treatment is    at least 15 consecutive days.-   17. The method of any one of embodiments 1-16, wherein the Compound    1 is administered to the patient once every 12 hours on consecutive    days throughout a course of treatment.-   18. The method of any one of embodiments 1-17, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 4 months.-   19. The method of any one of embodiments 1-17, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 6 months.-   20. A method of treating a patient diagnosed with a cancer selected    from the group consisting of: glioma, chondrosarcoma, hepatobiliary,    and intrahepatic cholangiocarcinoma, the cancer harboring an IDH1    R132 mutation, the method comprising administering to the patient in    need thereof a therapeutically effective amount of a mIDH1 Inhibitor    Therapy throughout a Course of Treatment of at least one 28-day    treatment cycle, the mIDH1 Inhibitor Therapy consisting of    -   a. Compound 1 in combination with azacitidine for the patient        diagnosed with glioma or chondrosarcoma cancer; or    -   b. Compound 1 in combination with a PD-1 inhibitor for the        patient diagnosed with hepatobiliary cancer; or    -   c. Compound 1 in combination with gemcitabine and cisplatin        chemotherapy for the patient diagnosed with intrahepatic        cholangiocarcinoma.-   21. The method of embodiment 20, wherein Compound 1 is administered    at a dose of 150 mg BID.-   22. The method of any one of embodiments 20-21, wherein the PD-1    inhibitor is nivolumab.-   23. The method of any one of embodiments 20-22, wherein the    nivolumab is administered at a dose of 240 mg every 2 weeks or 480    mg every 4 weeks.-   24. The method of any one of embodiments 20-23, wherein the    azacitidine is administered at a dose of 75 mg/m², SC, d1-7, q4 wk    throughout the Course of Treatment.-   25. The method of any one of embodiments 20-24, wherein the Course    of Treatment is at least 4 months.-   26. The method of any one of embodiments 20-25, wherein the Course    of Treatment is at least 6 months.-   27. The method of any one of embodiments 1-26, wherein the Compound    1 is administered as an oral dosage form obtained from Example 1.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a chondrosarcoma    cancer harboring a cancer cell with an IDH1 R132 mutation, the    method comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 in combination with a    therapeutically effective amount of azacitidine.-   2. The method of embodiment 1, wherein Compound 1 is administered at    a dose of 150 mg taken orally twice daily.-   3. The method of any one of embodiments 1-2, wherein Compound 1 is    orally administered as the solid form obtained from Example 1.-   4. The method of any one of embodiments 1-3, further comprising the    steps of:    -   a. selecting a patient diagnosed with the cancer harboring an        IDH1 mutation;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        for a treatment cycle of 28 consecutive days.-   5. The method of any one of embodiments 1-4, wherein the cancer does    not harbor a IDH2 mutation.-   6. The method of any one of embodiments 1-5, wherein the cancer does    not harbor a IDH2 mutation selected from the group consisting of:    IDH2 R172K and IDH2 R140Q.-   7. The method of any one of embodiments 1-6, comprising the step of    detecting the IDH1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   8. The method of any one of embodiments 1-7, wherein administration    of Compound 1 to the patient results in a decreased    2-hydroxyglutarate (2-HG) levels in the blood of the patient after    the first 15 consecutive days of treatment of the patient with    Compound 1.-   9. The method of any one of embodiments 1-8, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   10. The method of any one of embodiments 1-9, wherein the method    comprises administering 150 mg of Compound 1 to the patient twice    daily throughout a course of treatment.-   11. The method of embodiment 10, wherein the course of treatment is    at least 15 consecutive days.-   12. The method of any one of embodiments 1-11, wherein the Compound    1 is administered to the patient once every 12 hours on consecutive    days throughout a course of treatment.-   13. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 4 months.-   14. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 6 months.-   15. A method of treating a patient diagnosed with a cancer, wherein    the cancer is chondrosarcoma, the cancer harboring an IDH1 R132    mutation, the method comprising administering to the patient in need    thereof a therapeutically effective amount of a mIDH1 Inhibitor    Therapy throughout a Course of Treatment of at least one 28-day    treatment cycle, the mIDH1 Inhibitor Therapy consisting of Compound    1 in combination with azacitidine for the patient diagnosed with    glioma or chondrosarcoma cancer.-   16. The method of embodiment 15, wherein Compound 1 is administered    at a dose of 150 mg BID.-   17. The method of any one of embodiments 15-16, wherein the    azacitidine is administered at a dose of 75 mg/m², SC, d1-7, q4 wk    throughout the Course of Treatment.-   18. The method of any one of embodiments 15-17, wherein the Course    of Treatment is at least 4 months.-   19. The method of any one of embodiments 15-18, wherein the Course    of Treatment is at least 6 months.-   20. The method of any one of embodiments 1-19, wherein the Compound    1 is administered as an oral dosage form obtained from Example 1.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with an intrahepatic    cholangiocarcinoma cancer harboring a cancer cell with an IDH1 R132    mutation, the method comprising administering to the patient in need    thereof a therapeutically effective amount of Compound 1 in    combination with a therapeutically effective amount of a gemcitabine    and cisplatin chemotherapy.-   2. The method of embodiment 1, wherein Compound 1 is administered at    a dose of 150 mg taken orally twice daily.-   3. The method of any one of embodiments 1-2, wherein Compound 1 is    orally administered as the solid form obtained from Example 1.-   4. The method of any one of embodiments 1-3, further comprising the    steps of:    -   a. selecting a patient diagnosed with the cancer harboring an        IDH1 mutation;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        for a treatment cycle of 28 consecutive days.-   5. The method of any one of embodiments 1-4, wherein the cancer does    not harbor a IDH2 mutation.-   6. The method of any one of embodiments 1-5, wherein the cancer does    not harbor a IDH2 mutation selected from the group consisting of:    IDH2 R172K and IDH2 R140Q.-   7. The method of any one of embodiments 1-6, comprising the step of    detecting the IDH1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   8. The method of any one of embodiments 1-7, wherein administration    of Compound 1 to the patient results in a decreased    2-hydroxyglutarate (2-HG) levels in the blood of the patient after    the first 15 consecutive days of treatment of the patient with    Compound 1.-   9. The method of any one of embodiments 1-8, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   10. The method of any one of embodiments 1-9, wherein the method    comprises administering 150 mg of Compound 1 to the patient twice    daily throughout a course of treatment.-   11. The method of embodiment 10, wherein the course of treatment is    at least 15 consecutive days.-   12. The method of any one of embodiments 1-11, wherein the Compound    1 is administered to the patient once every 12 hours on consecutive    days throughout a course of treatment.-   13. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 4 months.-   14. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 6 months.-   15. A method of treating a patient diagnosed with a cancer, wherein    the cancer is intrahepatic cholangiocarcinoma, the cancer harboring    an IDH1 R132 mutation, the method comprising administering to the    patient in need thereof a therapeutically effective amount of a    mIDH1 Inhibitor Therapy throughout a Course of Treatment of at least    one 28-day treatment cycle, the mIDH1 Inhibitor Therapy consisting    of Compound 1 in combination with gemcitabine and cisplatin    chemotherapy for the patient diagnosed with intrahepatic    cholangiocarcinoma.-   16. The method of embodiment 15, wherein Compound 1 is administered    at a dose of 150 mg BID.-   17. The method of any one of embodiments 15-16, wherein the Course    of Treatment is at least 4 months.-   18. The method of any one of embodiments 15-17, wherein the Course    of Treatment is at least 6 months.-   19. The method of any one of embodiments 1-18, wherein the Compound    1 is administered as an oral dosage form obtained from Example 1.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a hepatobiliary    cancer harboring a cancer cell with an IDH1 R132 mutation, the    method comprising administering to the patient in need thereof a    therapeutically effective amount of Compound 1 in combination with a    therapeutically effective amount of a PD-1 inhibitor.-   2. The method of embodiment1, wherein Compound 1 is administered at    a dose of 150 mg taken orally twice daily.-   3. The method of any one of embodiments 1-2, wherein Compound 1 is    orally administered as the solid form obtained from Example 1.-   4. The method of any one of embodiments 1-3, further comprising the    steps of:    -   a. selecting a patient diagnosed with the cancer harboring an        IDH1 mutation;    -   b. administering Compound 1 to the selected patient from        step (a) at a starting dose of 150 mg taken orally twice daily        for a treatment cycle of 28 consecutive days.-   5. The method of any one of embodiments 1-4, wherein the cancer does    not harbor a IDH2 mutation.-   6. The method of any one of embodiments 1-5, wherein the cancer does    not harbor a IDH2 mutation selected from the group consisting of:    IDH2 R172K and IDH2 R140Q.-   7. The method of any one of embodiments 1-6, comprising the step of    detecting the IDH1 mutation in a cell from the patient using a    next-generation sequencing (NGS)-based tumor genotyping assay.-   8. The method of any one of embodiments 1-7, wherein administration    of Compound 1 to the patient results in a decreased    2-hydroxyglutarate (2-HG) levels in the blood of the patient after    the first 15 consecutive days of treatment of the patient with    Compound 1.-   9. The method of any one of embodiments 1-8, wherein the method    comprises administering 150 mg of Compound 1 to the patient in the    solid form obtained from the method of Example 1.-   10. The method of any one of embodiments 1-9, wherein the method    comprises administering 150 mg of Compound 1 to the patient twice    daily throughout a course of treatment.-   11. The method of embodiment 10, wherein the course of treatment is    at least 15 consecutive days.-   12. The method of any one of embodiments 1-11, wherein the Compound    1 is administered to the patient once every 12 hours on consecutive    days throughout a course of treatment.-   13. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 4 months.-   14. The method of any one of embodiments 1-12, wherein the Compound    1 is administered to the patient throughout a course of treatment of    at least 6 months.-   15. A method of treating a patient diagnosed with a cancer, wherein    the cancer is hepatobiliary cancer, the cancer harboring an IDH1    R132 mutation, the method comprising administering to the patient in    need thereof a therapeutically effective amount of a mIDH1 Inhibitor    Therapy throughout a Course of Treatment of at least one 28-day    treatment cycle, the mIDH1 Inhibitor Therapy consisting of Compound    1 in combination with a PD-1 inhibitor for the patient diagnosed    with hepatobiliary cancer.-   16. The method of embodiment 15, wherein Compound 1 is administered    at a dose of 150 mg BID.-   17. The method of any one of embodiments 15-16, wherein the PD-1    inhibitor is nivolumab.-   18. The method of any one of embodiments 15-17, wherein the    nivolumab is administered at a dose of 240 mg every 2 weeks or 480    mg every 4 weeks.-   19. The method of any one of embodiments 15-18, wherein the Course    of Treatment is at least 4 months.-   20. The method of any one of embodiments 15-19, wherein the Course    of Treatment is at least 6 months.-   21. The method of any one of embodiments 1-20, wherein the Compound    1 is administered as an oral dosage form obtained from Example 1.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. Use of a pharmaceutical composition comprising Compound 1, or    pharmaceutically acceptable salt thereof,

-   -   in treating a cancer harboring an isocitrate dehydrogenase-1        (IDH-1) mutation (mIDH-1) in a patient by administering a total        of 300 mg of Compound 1 (or a corresponding amount in the form        of a pharmaceutically acceptable salt thereof) to a patient each        day during a course of treatment.

-   2. The use of embodiment 1, wherein a 150 mg amount of Compound 1(or    a corresponding amount in the form of a pharmaceutically acceptable    salt thereof) is administered to the patient twice per day (BID)    throughout the course of treatment.

-   3. The use of embodiment 1, wherein the cancer is a mIDH-1 form of    acute myeloid leukemia.

-   4. The use of embodiment 3, wherein the acute myeloid leukemia is    relapsed or refractory or is drug-resistant.

-   5. The use of embodiment 1, wherein the cancer is a mIDH-1 solid    tumor.

-   6. The use of embodiment 1, wherein the cancer is of a mIDH-1    glioma.

-   7. The use of embodiment 6, wherein the mIDH-1 glioma is an advanced    glioma that has recurred or progressed prior to the administration    of Compound 1.

-   8. The use of any one of the preceding embodiments, wherein the    mIDH1 is a R132X mutation.

-   9. The use of claim 8, wherein the R132X mIDH-1 mutation is selected    from R132L, R132G and R132S.

-   10. The use of any one of the preceding embodiments, wherein the    pharmaceutical composition comprising Compound 1 or a    pharmaceutically acceptable salt thereof is orally administered to    the patient.

-   11. The use of any one of the preceding embodiments, wherein    Compound 1, (or a corresponding amount in the form of a    pharmaceutically acceptable salt thereof)is administered as a single    agent for the treatment of the cancer harboring the IDH-1 mutation.

-   12. The use of any of the preceding embodiments, wherein the course    of treatment is at least 15 consecutive days to reach a steady state    blood concentration of Compound 1 (or a corresponding amount in the    form of a pharmaceutically acceptable salt thereof)in the patient.

-   13. The use of any one of the preceding embodiments, wherein the    course of treatment is at least 6 months.

-   14. The use of any one of the preceding embodiments, wherein the    pharmaceutical composition comprises Compound 1 in a Type A solid    form characterized by a reflection X-ray powder diffraction (XRPD)    pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6,    and 27.8 degrees ±0.2°2θ.

-   15. The use of any one of the preceding embodiments, wherein the    pharmaceutical composition comprises the following formulation for    oral administration: (a) Type A solid form of Compound 1 in a    relative weight of about 33, (b) a microcrystalline cellulose in a    relative weight of about 61, (c) a croscamellose sodium in a    relative weight of about 5 and a magnesium stearate in a relative    weight of about 1;    -   wherein the Type A solid form of Compound 1 is characterized by        a reflection X-ray powder diffraction (XRPD) pattern comprising        characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees        ±0.2°2θ.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A pharmaceutical composition comprising Compound 1:

-   -   or pharmaceutically acceptable salt thereof, for use in treating        a patient diagnosed with a form of cancer harboring an        isocitrate dehydrogenase-1 (IDH-1) mutation (mIDH-1) by        administering a total of 300 mg of Compound 1 (or a        corresponding amount in the form of a pharmaceutically        acceptable salt thereof) to the patient each day during a course        of treatment.

-   2. The pharmaceutical composition of embodiment 1, wherein a 150 mg    amount of Compound 1 (or a corresponding amount in the form of a    pharmaceutically acceptable salt thereof) is administered to the    patient twice per day (BID) throughout the course of treatment.

-   3. The pharmaceutical composition of embodiment 1, wherein the    cancer is a mIDH-1 form of acute myeloid leukemia.

-   4. The pharmaceutical composition of embodiment 3, wherein the acute    myeloid leukemia is relapsed or refractory or is drug-resistant.

-   5. The pharmaceutical composition of embodiment 1, wherein the    cancer is a mIDH-1 solid tumor.

-   6. The pharmaceutical composition of embodiment 1, wherein the    cancer is of a mIDH-1 glioma.

-   7. The pharmaceutical composition of embodiment 6, wherein the    mIDH-1 glioma is an advanced glioma that has recurred or progressed    prior to the administration of Compound 1.

-   8. The pharmaceutical composition of any one of the preceding    embodiments, wherein the mIDH1 is a R132X mutation.

-   9. The pharmaceutical composition of embodiment 8, wherein the R132X    mIDH-1 mutation is selected from R132L, R132G and R132S.

-   10. The pharmaceutical composition of any one of the preceding    embodiments, wherein the pharmaceutical composition comprising    Compound 1 or a pharmaceutically acceptable salt thereof is orally    administered to the patient.

-   11. The pharmaceutical composition of any one of the preceding    embodiments, wherein Compound 1, (or a corresponding amount in the    form of a pharmaceutically acceptable salt thereof) is administered    as a single agent for the treatment of the cancer harboring the    IDH-1 mutation.

-   12. The pharmaceutical composition of any of the preceding    embodiments, wherein the course of treatment is at least 15    consecutive days to reach a steady state blood concentration of    Compound 1 (or a corresponding amount in the form of a    pharmaceutically acceptable salt thereof) in the patient.

-   13. The pharmaceutical composition of any one of the preceding    embodiments, wherein the course of treatment is at least 6 months.

-   14. The pharmaceutical composition of any one of the preceding    embodiments, wherein the pharmaceutical composition comprises    Compound 1 in a Type A solid form characterized by a reflection    X-ray powder diffraction (XRPD) pattern comprising characteristic    peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2°2θ.

-   15. The pharmaceutical composition of any one of the preceding    embodiments, wherein the pharmaceutical composition comprises the    following formulation for oral administration: (a) Type A solid form    of Compound 1 in a relative weight of about 33, (b) a    microcrystalline cellulose in a relative weight of about 61, (c) a    croscamellose sodium in a relative weight of about 5 and a magnesium    stearate in a relative weight of about 1; wherein the Type A solid    form of Compound 1 is characterized by a reflection X-ray powder    diffraction (XRPD) pattern comprising characteristic peaks at 6.3,    12.8, 13.8, 23.6, and 27.8 degrees ±0.2°2θ.

The present disclosure also contemplates, among other things, thefollowing numbered embodiments:

-   1. A method of treating a patient diagnosed with a form of cancer    harboring a R132X IDH1 mutation, the method comprising administering    to the patient in need thereof a total of 150 mg of the compound of    Formula (1) twice per day (BID)

-   2. The method of embodiment 1 above, wherein the R132X IDH1 mutation    is selected from the group consisting of: R132H and R132C IDH1    mutations.-   3. The method of embodiment 1 above, wherein the R132X IDH1 mutation    is selected from the group consisting of: R132S, R132G, and R132L    IDH1 mutations.-   4. The method of any one of the enumerated embodiments above,    wherein the compound of Formula (1) is orally administered to the    patient.-   5. The method of any one of the enumerated embodiments above,    further comprising administering a total of 150 mg BID of the    compound of Formula (1) to the patient throughout a course of    treatment of at least 15 consecutive days.-   6. The method of embodiment 5 above, further comprising    administering a total of 150 mg BID of the compound of Formula (1)    to the patient on consecutive days throughout a course of treatment    of between 15 days and 6 months.-   7. The method of embodiment 5 above, further comprising    administering a total of 150 mg BID of the compound of Formula (1)    to the patient on consecutive days throughout a 6 month course of    treatment.-   8. The method of any one of the enumerated embodiments 1-7 above,    wherein the compound of Formula (1) is administered to the patient    in an oral unit dosage form.-   9. A method of treating a patient diagnosed with acute myeloid    leukemia (AML) having an IDH1 mutation, the method comprising    administering to the patient in need thereof a combination of    azacitidine and the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   10. The method of embodiment 9 above, wherein the acute myeloid    leukemia is relapsed or refractory or is drug-resistant.

-   11. The method of any one of the embodiments 9-10 above, wherein the    IDH1 mutation is a R132X IDH1 mutation.

-   12. The method of any one of the embodiments 9-11 above, wherein the    R132X IDH1 mutation is selected from the group consisting of: R132H    and R132C IDH1 mutations.

-   13. The method of any one of the embodiments 9-11 above, wherein the    R132X IDH1 mutation is selected from the group consisting of: R132S,    R132G, and R132L IDH1 mutations.

-   14. The method of any one of the embodiments 9-13 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   15. The method of any one of the embodiments 9-13 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   16. The method of any one of the embodiments 9-13 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   17. A method of treating a patient diagnosed with myelodysplastic    syndrome (MDS) having an IDH1 mutation, the method comprising    administering to the patient in need thereof a combination of    azacitidine and the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   18. The method of embodiment 17 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   19. The method of embodiment 18 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   20. The method of embodiment 18 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   21. The method of any one of the embodiments 17-20 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   22. The method of any one of the embodiments 17-20 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   23. The method of any one of the embodiments 17-20 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   24. A method of treating a patient diagnosed with myelodysplastic    syndrome (MDS) having an IDH1 mutation, the method comprising    administering to the patient in need thereof the compound of Formula    (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day.

-   25. The method of embodiment 24 above, wherein the acute myeloid    leukemia is relapsed or refractory or is drug-resistant.

-   26. The method of any one of the embodiments 24-25 above, wherein    the IDH1 mutation is a R132X IDH1 mutation.

-   27. The method of embodiment 26 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   28. The method of embodiment 26 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   29. The method of any one of the embodiments 24-28 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   30. The method of any one of the embodiments 24-28 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   31. The method of any one of the embodiments 24-28 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   32. A method of treating a patient diagnosed with glioma having an    IDH1 mutation, wherein the method comprises administering to the    patient in need thereof the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day.

-   33. The method of any one of the embodiments 32 above, wherein the    IDH1 mutation is a R132X IDH1 mutation.

-   34. The method of embodiment 33 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   35. The method of embodiment 34 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   36. The method of any one of the embodiments 32-35 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   37. The method of any one of the embodiments 32-35 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   38. The method of any one of the embodiments 32-35 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   39. A method of treating a patient diagnosed with chondrosarcoma    having an IDH1 mutation, wherein the method comprises administering    to the patient in need thereof the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day.

-   40. The method of any one of the embodiments 39 above, wherein the    IDH1 mutation is a R132X IDH1 mutation.

-   41. The method of embodiment 40 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   42. The method of embodiment 40 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   43. The method of any one of the embodiments 39-42 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   44. The method of any one of the embodiments 39-42 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   45. The method of any one of the embodiments 39-42 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   46. A method of treating a patient diagnosed with hepatobiliary    cholangiocarcinoma having an IDH1 mutation, wherein the method    comprises administering to the patient in need thereof the compound    of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day.

-   47. The method of embodiment 46 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   48. The method of embodiment 47 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   49. The method of embodiment 47 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   50. The method of any one of the embodiments 46-49 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   51. The method of any one of the embodiments 46-49 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   52. The method of any one of the embodiments 46-49 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   53. A method of treating a patient diagnosed with intrahepatic    cholangiocarcinoma having an IDH1 mutation, wherein the method    comprises administering to the patient in need thereof the compound    of Formula (1).

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day.

-   54. The method of embodiment 53 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   55. The method of embodiment 54 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   56. The method of embodiment 54 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   57. The method of any one of the embodiments 53-56 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   58. The method of any one of the embodiments 53-56 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   59. The method of any one of the embodiments 53-56 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   60. A method of treating a patient diagnosed with glioma having an    IDH1 mutation, wherein the method comprises administering to the    patient in need thereof a combination of azacitidine and the    compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   61. The method of embodiment 60 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   62. The method of embodiment 61 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   63. The method of embodiment 61 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   64. The method of any one of the embodiments 60-63 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   65. The method of any one of the embodiments 60-63 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   66. The method of any one of the embodiments 60-63 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   67. A method of treating a patient diagnosed with chondrosarcoma    having an IDH1 mutation, wherein the method comprises administering    to the patient in need thereof a combination of azacitidine and the    compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   68. The method of embodiment 67 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   69. The method of embodiment 68 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   70. The method of embodiment 68 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   71. The method of any one of the embodiments 67-70 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   72. The method of any one of the embodiments 67-70 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   73. The method of any one of the embodiments 67-70 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   74. A method of treating a patient diagnosed with hepatobiliary    cholangiocarcinoma having an IDH1 mutation, wherein the method    comprises administering to the patient in need thereof a combination    of azacitidine and the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   75. The method of embodiment 74 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   76. The method of embodiment 75 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   77. The method of embodiment 75 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   78. The method of any one of the embodiments 74-77 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   79. The method of any one of the embodiments 74-77 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   80. The method of any one of the embodiments 74-77 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   81. A method of treating a patient diagnosed with intrahepatic    cholangiocarcinoma having an IDH1 mutation, wherein the method    comprises administering to the patient in need thereof a combination    of azacitidine and the compound of Formula (1),

-   -   wherein a total of 150 mg of the compound of Formula (1) is        administered to the patient twice per day (BID) each day, and        the azacitidine is administered to the patient at a total dose        of 75 mg/m² each day for 7 consecutive days beginning at the        start of each treatment cycle, followed 21 consecutive days        without administration of the azacitidine to the patient.

-   82. The method of embodiment 81 above, wherein the IDH1 mutation is    a R132X IDH1 mutation.

-   83. The method of embodiment 82 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132H and R132C    IDH1 mutations.

-   84. The method of embodiment 82 above, wherein the R132X IDH1    mutation is selected from the group consisting of: R132S, R132G, and    R132L IDH1 mutations.

-   85. The method of any one of the embodiments 81-84 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of between 15 days and 6 months.

-   86. The method of any one of the embodiments 81-84 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 15 days.

-   87. The method of any one of the embodiments 81-84 above, further    comprising administering a total of 150 mg BID of the compound of    Formula (1) to the patient on consecutive days throughout a course    of treatment of at least 6 months.

-   88. The method of any one of the embodiments 1-87 above, wherein the    compound of Formula (1) is administered in a crystalline form.

-   89. The method of any one of the embodiments 1-88 above, wherein the    compound of Formula (1) is administered as a Type A crystalline    form.

-   90. The method of any one of the embodiments 1-89 above, wherein the    compound of Formula (1) is orally administered in a capsule    comprising a total of 150 mg of the compound of Formula (1).

-   91. The method of any one of the embodiments 1-90 above, wherein the    compound of Formula (1) is orally administered in multiple capsules    each comprising a total of 50 mg of the compound of Formula (1).

The present disclosure enables one of skill in the relevant art to makeand use the inventions provided herein in accordance with multiple andvaried embodiments. Various alterations, modifications, and improvementsof the present disclosure that readily occur to those skilled in theart, including certain alterations, modifications, substitutions, andimprovements are also part of this disclosure. Accordingly, theforegoing description are by way of example to illustrate thediscoveries provided herein.

EXAMPLES Example 1 Synthesis of(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile(Compound 1)

Compound 1 can be prepared in a convergent synthesis from Intermediate Aand Intermediate B as shown in FIG. 23 via the nucleophilic displacementreaction under basic conditions of(S)-3-(1-aminoethyl)-6-chloroquinolin-2(1H)-one (Intermediate A) and thefluoropyridone (Intermediate B). ¹H, ¹³C NMR and mass spectral data areconsistent with the assigned structure. The asymmetric synthesis ofIntermediate A started with the condensation of the commerciallyavailable quinoline aldehyde (1) with (R)-tert-butanesulfinamide (2) toform the chiral (R)-N-tert-butanesulfinimine (3), followed by additionof methyl magnesium bromide in dichloromethane to yield the desiredproduct (4) as the major diastereoisomer (dr: 98:2). Cleavage of thechiral auxiliary and simultaneous hydrolysis of 2-chloroquinoline moietyunder mildly acidic conditions using 1N HCl in dioxane gave IntermediateA in quantitative yield. The structure of Intermediate A was confirmedby NMR and mass spectroscopy, and the enantiomeric purity was determinedby chiral SFC analysis. The (S)-stereochemistry was confirmed by X-rayco-crystal structures of several inhibitor analogs prepared from thesame chiral amine intermediate bound to mIDH-1 R132H. Intermediate (B)was prepared from commercially available 5-fluoropicolinonitrile in foursteps. N-oxidation of 5-fluoropicolinonitrile followed by reflux of theN-oxide in acetic anhydride gave the acetate, following work-up andpurification. Solvolysis of the acetate group followed by N-methylationunder standard conditions gave a mixture of N-methylated andO-methylated products (4:1). The minor O-methylated product was removedby column chromatography. NMR and mass spectral data are consistent withthe structure of Intermediate Compound (B). Compound 1(5-{[(1S)-1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl]amino}-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile)has a molecular weight of 355 with a melting point onset temperature of251.3° C. (DSC) and peak maximum 254.1° C.

Intermediate 1: (S)-3-(1-aminoethyl)-6-chloroquinolin-2(1H)-onehydrochloride

Step-1:(R,E)-N-((2,6-dichloroquinolin-3-yl)methylene)-2-methylpropane-2-sulfinamide

To a mixture of 2,6-dichloroquinoline-3-carbaldehyde (15.0 g, 66.37mmol) and (R)-2-methylpropane-2-sulfinamide (8.85 g, 73.14 mmol) in1,2-dichloroethane (150 mL) was added CuSO₄ (16.0 g, 100.25 mmol). Theresulting mixture was heated to 55° C. and stirred at 55° C. overnight.After TLC and MS showed complete disappearance of starting materials,the mixture was cooled to room temperature and filtered through a pad ofCelite®. The pad of Celite® was then rinsed with CH₂Cl₂. The filtratewas evaporated to dryness in vacuo and purified by SiO₂ columnchromatography (0 to 25% hexanes/EtOAc) to afford the title compound,(R,E)-N-((2,6-dichloroquinolin-3-yl)methylene)-2-methylpropane-2-sulfinamide,as a yellow solid (17.7 g, 81% yield).

Step-2:(R)-N-((S)-1-(2,6-dichloroquinolin-3-yl)ethyl)-2-methylpropane-2-sulfinamide

To a solution of(R,E)-N-((2,6-dichloroquinolin-3-yl)methylene)-2-methylpropane-2-sulfinamide(8.85 g, 26.88 mmol) in anhydrous CH₂Cl₂ (200 mL) at −60° C. was addeddropwise MeMgBr (3M solution in diethyl ether, 13.5 mL, 40.54 mmol). Theresulting reaction mixture was stirred at about −60 to −50° C. for 3hours and then stirred at −20° C. overnight under an atmosphere of N₂.After TLC and MS showed complete disappearance of starting materials,saturated NH₄C1 (163 mL) was added at −20° C. and the resulting mixturewas stirred for 10 minutes. The aqueous phase was extracted with CH₂Cl₂(100 mL×3), dried over anhydrous Na₂SO₄, filtered, and evaporated. Theresidue was purified by column chromatography on an ISCO® chromatographysystem (SiO₂: Gold column; gradient; hexanes to 100% EtOAc) to providethe title compound,(R)-N-((S)-1-(2,6-dichloroquinolin-3-yl)ethyl)-2-methylpropane-2-sulfinamide,as a yellow solid (5.8 g, 63% yield).

Step-3: (S)-3-(1-aminoethyl)-6-chloroquinolin-2(1H)-one hydrochloride(A)

A mixture of(R)-N-((S)-1-(2,6-dichloroquinolin-3-yl)ethyl)-2-methylpropane-2-sulfinamide(6.6 g, 19.13 mmol) in 1,4-dioxane (41 mL) and 1N HCl (41 mL) was heatedat reflux overnight. The solvents were evaporated in vacuo and theresulting residue was dissolved in hot water and lyophilized. The crudeproduct was triturated with diethyl ether to afford the title compound Aas a yellow solid (9.0 g, ee: 98.4%). ¹H NMR (300 MHz, DMSO-d₆): δ ppm12.4 (br s, 1H), 8.32 (br s, 2H), 8.07 (s, 1H), 7.85 (d, J=2.2 Hz, 1H),7.63 (dd, J1=8.8 Hz, J2=2.5 Hz, 1H), 7.40 (d, J=8.8 Hz, 1H), 4.40-4.45(m, 1H), 1.53(d, J=8.5 Hz, 3H). LCMS (Method 2): Rt 3.42 min, m/z 223.1[M+H]⁺.

Intermediate 2:5-fluoro-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile

Step-1: 2-cyano-5-fluoropyridine 1-oxide

A solution of 5-fluoropicolinonitrile (7.27 g, 59.5 mmol) in CHCl₃ (60mL) was added dropwise by addition funnel to a solution of m-CPBA (<77%,22.00 g, 98 mmol) in CHCl₃ 160 mL). The solution was stirred at refluxfor 4 days, at which time LCMS showed ˜85% conversion. The sample wasallowed to cool, then sodium sulfite (12.4 g, 98 mmol) was added and thesample was stirred at room temperature three hours, during which timethe solution became thick with a white precipitate. The sample wasdiluted with DCM (300 mL) and filtered on a Buchner funnel, and thefilter cake was washed with DCM (˜400 mL). A white material precipitatedin the filtrate. The filtrate mixture was washed with saturated aqueousNaHCO₃ (400 mL), during which the solids went into solution. The organiclayer was washed with water (300 mL), then dried (MgSO₄) and filtered.Silica gel was added and the mixture was evaporated under reducedpressure. The material was chromatographed by Biotage MPLC (340 g silicagel column) with 0 to 100% EtOAc in hexanes, with isocratic elution whenpeaks came off to provide 2-cyano-5-fluoropyridine 1-oxide (4.28 g, 31.0mmol, 52% yield) as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm8.85-8.93 (m, 1H), 8.23 (dd, J=9.09, 6.74 Hz, 1H), 7.53-7.64 (m, 1H).LCMS (Method 1): Rt 0.57 min., m/z 138.9 [M+H]⁺.

Step 2: 6-cyano-3-fluoropyridin-2-yl acetate

A solution of 2-cyano-5-fluoropyridine 1-oxide (4.28 g, 31.0 mmol) inacetic anhydride (40 ml, 424 mmol) was heated at reflux (150° C. bath)three days, during which the clear solution turned dark. The sample wasconcentrated under reduced pressure. The residue was dissolved in MeOH(30 mL) and stirred 1 hour. Silica gel was added and the solvent wasevaporated under reduced pressure. The material was chromatographed byBiotage MPLC (100 g silica gel column) with 0 to 23% EtOAc in hexanes toprovide 6-cyano-3-fluoropyridin-2-yl acetate (3.32 g, 18.43 mmol, 60%yield) as a clear liquid that solidified on cooling. ¹H NMR (300 MHz,CHLOROFORM-d): δ ppm 7.65-7.75 (m, 2H), 2.42 (s, 3H). LCMS (Method 1):Rt 1.54 min., m/z 138.8 (loss of acetate).

Step 3: 5-fluoro-6-oxo-1,6-dihydropyridine-2-carbonitrile

A solution of 6-cyano-3-fluoropyridin-2-yl acetate (3.32 g, 18.43 mmol)in MeOH (40 ml) was treated with potassium carbonate (5.10 g, 36.9 mmol)and stirred at room temperature for four hours. LCMS at 2 hours showedthe reaction had gone to completion. The solvent was evaporated underreduced pressure. The residue was dissolved in water (100 mL) andacidified to pH with 1M HCl. The solution was extracted with EtOAc(3×100 mL). The combined organic extracts were dried (Na₂SO₄), filtered,and evaporated under reduced pressure to provide5-fluoro-6-oxo-1,6-dihydropyridine-2-carbonitrile (2.34 g, 16.94 mmol,92% yield) as a white solid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 12.92 (brs, 1H), 7.73 (br s, 1H), 7.43 (br s, 1H). LCMS (Method 1): Rt 0.70 min.,m/z 138.9 [M+H]⁺.

Step 4: 5-fluoro-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile (B)

A mixture of 5-fluoro-6-oxo-1,6-dihydropyridine-2-carbonitrile (2.31 g,16.73 mmol) and potassium carbonate (4.86 g, 35.2 mmol) in a 200 mLround bottom flask was treated with DMF (46 mL) and stirred 15 minutes.MeI (1.2 mL, 19.19 mmol) was added and the mixture was stirred at roomtemperature 45 minutes. The solvent was evaporated under reducedpressure. The residue was mixed with water (150 mL) and extracted withDCM (2×150 mL). The combined organic extracts were dried (MgSO₄),filtered, treated with silica gel, and evaporated under reducedpressure, then evaporated further at 60° C. under high vacuum. Thematerial was chromatographed by Biotage MPLC with 0 to 35% EtOAc inhexanes, with isocratic elution at 16% EtOAc and 35% EtOAc while peakscame off. The peak that came off with 16% EtOAc was 0-methylatedmaterial and was discarded. The peak that came off with 35% EtOAcprovided the title compound B (1.70 g, 11.17 mmol, 67% yield) as asolid. ¹H NMR (300 MHz, DMSO-d₆): δ ppm 7.53 (dd, J=9.38, 7.62 Hz, 1H),7.18 (dd, J=7.77, 4.84 Hz, 1H), 3.60 (s, 3H). LCMS (Method 1): Rt 0.94min., m/z 152.9 [M+H]⁺.

Step 5:(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile(Compound 1)

A mixture of 5-fluoro-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrileB (1.23 g, 8.09 mmol), (S)-3-(1-aminoethyl)-6-chloroquinolin-2(1H)-onehydrochloride A (1.91 g, 7.37 mmol) and N,N-diisopropylethylamine (3.8mL, 21.8 mmol) in anhydrous dimethyl sulfoxide (57 mL) under N₂ washeated to 110° C. and stirred for 6 hours. After cooling to roomtemperature, the mixture was partitioned between EtOAc/H₂O (750 mL/750mL). The organic layer was separated, dried (Na₂SO₄) and concentrated invacuum. The residue was purified on ISCO twice (40 g silica gel column,EtOAc/hexanes 0˜100%; 80 g silica gel column, MeOH/dichloromethane0˜5%). The colorless fractions were combined and dichloromethane wasremoved under reduced pressure on rotavap until a lot of white solidprecipitated out. The white solid was collected by filtration and washedwith cold MeOH. It was then mixed with MeCN/H₂O (10 mL/25 mL) andlyophilized to afford the title Compound 1 as a white solid (790 mg).m.p. 262-264° C. ¹H NMR (300 MHz, DMSO-d₆) δ: 12.07 (s, 1H), 7.75 (s,1H), 7.73 (d, J=2.2 Hz, 1H), 7.51 (dd, J=8.6, 2.3 Hz, 1H), 7.31 (d,J=8.8 Hz, 1H), 6.97 (d, J=8.0 Hz, 1H), 6.93 (d, J=7.7 Hz, 1H), 5.95 (d,J=8.0 Hz, 1H), 4.68 (m, 1H), 3.58 (s, 3H), 1.50 (d, J=6.6 Hz, 3H). LCMS(Method 2): 100% pure @ 254 nm, Rt 10.78 min, m/z 355, 357 [M+H]+. Thefiltrate and the colored fractions (TLC pure) from the second ISCO werecombined and treated with activated charcoal and filtered (until thefiltrate is colorless). The filtrate was then concentrated under reducedpressure on rotavap to remove dichloromethane until a lot of white solidprecipitated out. The white solid was collected by filtration and washedwith cold MeOH. It was then mixed with MeCN/H₂O (10 mL/25 mL) andlyophilized to afford the title Compound 1 as a white solid (970 mg).m.p. 262-264° C. ¹H NMR (300 MHz, DMSO-d₆) δ: 12.06 (s, 1H), 7.75 (s,1H), 7.73 (d, J=2.5 Hz, 1H), 7.51 (dd, J=8.6, 2.3 Hz, 1H), 7.31 (d,J=8.8 Hz, 1H), 6.97 (d, J=8.0 Hz, 1H), 6.92 (d, J=8.0 Hz, 1H), 5.95 (d,J=8.0 Hz, 1H), 4.68 (m, 1H), 3.58 (s, 3H), 1.50 (d, J=6.9 Hz, 3H). LCMS(Method 2): 100% pure @254 nm, m/z 355, 357 [M+H]+. The total yield forcombined two batches is 67%.

Step 6: Solid Form of(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile

Unless otherwise indicated, the clinical trial in Examples 10-13 wereperformed using a pharmaceutically acceptable solid form in an oraldosage form of Compound 1 that can be obtained by the method of Step 6of Example 1. All volumes are with respect to the quantity of Compound 1(v/w). Compound 1 is dissolved in 18 volumes of dichloromethane. Theresulting solution is then concentrated under reduced pressure toapproximately 5 volumes. To the mixture is added 5 volumes of ethylacetate. The mixture is concentrated under reduced pressure to 5volumes. To the mixture is added an additional 5 volumes of ethylacetate, and the mixture again concentrated under reduced pressure to 5volumes. The mixture is diluted to 10 volumes with ethyl acetate, andthe mixture stirred at room temperature for 18 hours and then cooled to0° C. The mixture is stirred at 0° C. for 3 hours and then filtered. Thesolids are rinsed with ethyl acetate and dried under vacuum(counterbalanced by nitrogen) at ambient temperature.

The crystalline solid was determined to be the solid form of Compound 1Type A. The DVS Isotherm of Compound 1 Type A is shown in FIG. 24. DVSshows maximum water uptake of 0.25% w/w at 25° C./90% RH, indicatingCompound 1 Type A is not hygroscopic. The thermal behavior Compound 1Type A was evaluated using DSC. An endothermic event was observed at256.6° C. (peak max). The onset temperature and heat of fusion were255.0° C. and 108.7 J/g respectively (FIG. 25). TGA data (FIG. 26) donot show significant release of moisture or nonaqueous residualvolatiles from Compound 1 Type A.

The X-ray powder diffraction pattern of the crystalline Compound 1 TypeA is depicted in FIG. 27, and the corresponding data is summarized inTable 9 below.

TABLE 9 2 theta ± d-spacing 0.2 Å ± 0.2 5.7 15.4 6.3 14.0 8.5 10.4 10.68.4 11.4 7.8 12.8 6.9 13.8 6.4 14.2 6.2 15.2 5.8 15.6 5.7 17.3 5.1 17.95.0 18.2 4.9 18.9 4.7 19.6 4.5 20.6 4.3 21.5 4.1 22.0 4.0 22.8 3.9 23.63.8 24.5 3.6 24.8 3.6 25.3 3.5 25.6 3.5 26.0 3.4 26.3 3.4 27.0 3.3 27.83.2 28.9 3.1 30.0 3.0 31.2 3.0 32.1 2.8 33.6 2.7 34.1 2.6 36.3 2.5 37.02.4 38.1 2.4

Preferably, the oral dosage form of Compound 1 is a solid formdesignated Type A that is characterized by a reflection X-ray powderdiffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8,13.8, 23.6, and 27.8 degrees ±0.2°2θ. High resolution X-ray PowderDiffraction experiments were performed with Panalytical X' Pert3 PowderXRPD on a Si zero-background holder. The 2 theta position was calibratedagainst Panalytical 640 Si powder standard. Details of the XRPD methodare listed below, with XRPD peaks reported as diffraction angles at 2theta, with d-spacing measured in angstroms.

Parameters for Reflection Mode X-Ray wavelength Cu, kα, Kα1 (A):1.540598, Kα2 A): 1.544426 Kα2/Kα1 intensity ratio: 0.50 X-Ray tubesetting 45 kV, 40 mA Divergence slit Automatic Scan mode Continuous Scanrange (°2TH) 3°-40° Step size (°2TH) 0.0131 Scan speed (°/g) 0.033 

Compound 1 is preferably administered in an oral unit dosage formcomprising a pharmaceutical composition that includes the followingformulation: (a) Type A solid form of Compound 1 (e.g., in a relativeweight of about 33), (b) a microcrystalline cellulose (e.g., in arelative weight of about 61), (c) a croscamellose sodium (e.g., in arelative weight of about 5) and a magnesium stearate (e.g., in arelative weight of about 1). The pharmaceutical composition for oraladministration can comprise Compound 1 (e.g. in a Type A solid form)with pharmaceutically acceptable excipients in a capsule or tablet. Forexample, a capsule may contain a total of 50 mg or 150 mg of Compound 1in a unit dosage form. The capsule may encapsulate the pharmaceuticalcomposition comprising Compound 1 in a relative weight of about 30-50%by weight relative to the weight of the pharmaceutical composition. Inanother embodiment, a GMP manufacturing batch can comprise Compound 1,optionally provided in the Type A solid form.

In particular, the Compound 1 Type A solid form can be characterized byan X-ray Powder Diffraction (XRPD), having diffractions at angles (2theta±0.2) of 6.3, 12.8, 13.8, 23.6, and 27.8. In some embodiments, anovel Compound 1 Type A is characterized by an X-ray Powder Diffraction(XRPD), having diffractions at angles (2 theta ±0.2) of 6.3, 12.8, 13.8,23.6, and 27.8, corresponding to d-spacing (angstroms ±0.2) of 14.0,6.9, 6.4, 3.8, and 3.2, respectively. In some embodiments, Compound 1Type A can be identified by X-ray Powder Diffraction (XRPD), havingcharacteristic diffractions at angles (2 theta ±0.2) of 5.7, 6.3, 8.5,10.6, 12.8, 13.8, 17.3, 22.0, 22.8, 23.6, and 27.8. In some embodiments,Compound 1 Type A can be identified by X-ray Powder Diffraction (XRPD),having characteristic diffractions at angles (2 theta} 0.2) of 5.7, 6.3,8.5, 10.6, 12.8, 13.8, 17.3, 22.0, 22.8, 23.6, and 27.8, correspondingto d-spacing (angstroms ±0.2) of 15.4, 14.0, 8.4, 6.9, 6.4, 5.1, 4.0,3.9, 3.8, and 3.2, respectively.

In some embodiments, Compound 1 Type A solid form is characterized by adifferential scanning calorimetry (DSC) endotherm having a minima atabout 256.64° C. Differential Scanning Calorimetry (DSC) experimentswere performed on TA Q2000 DSC from TA Instruments. Samples were heatedat 10° C./min from about 20° C. to about 300° C. using dry nitrogen topurge the system. The details of the method are provided below:

Parameters DSC Pan Type Aluminum pan, closed Temperature RT-250° C. Ramprate 10° C./min Purge gas N₂

Step 7: Oral Solid Dosage Form of(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile

The oral dosage form of Compound 1 is a pharmaceutically acceptablesolid form of the compound(S)-5-((1-(6-chloro-2-oxo-1,2-dihydroquinolin-3-yl)ethyl)amino)-1-methyl-6-oxo-1,6-dihydropyridine-2-carbonitrile,can be obtained using the method of Example 1 Step 6. The oral dosageform does not contain associated solvent or a counter ion. Inparticular, the oral dosage form of Compound 1 can be a capsulecomprising drug substance (Compound 1) blended with excipients toimprove powder flow and encapsulated in a Coni-Snap® hard gelatincapsule suitable for oral dosage in humans.

Example 2 Compound 1 Potently and Selectively Inhibited 2-HG Productionin IDH1 R132H and IDH1 R132C Mutant Enzymes in Biochemical Assays,Compared to Wild Type IDH1 Enzyme and Mutant IDH2 Enzymes

The biochemical potencies of Compound 1 against IDH1 R132H and IDH1R132C mutants were determined in diaphorase-coupled assays, whichmeasure activity by the determination of the level of remainingco-substrate NADPH after the enzymatic reaction is quenched. Recombinanthomodimeric IDH1 R132H or IDH1 R132C mutant enzymes were used in theseassays.

In order to evaluate the cellular potency of Compound 1 for otherIDH1^(R132) mutations that have been identified in human cancers,IDH1^(R132L), IDH1^(R132G) and IDH1^(R132S) were expressed in U87MGhuman glioblastoma cells. Matched IDH1^(R132H) and IDH1^(R132C) lineswere also prepared to allow direct comparisons in the same cellularbackground, as well as to compare the effects observed from the samemutation in different cell lines. As for the HT1080 and HCT-116 celllines described above, the engineered mIDH1-expressing U87MG cellsproduced higher concentrations of 2-HG but exhibited a similar growthrate when compared to parental U87MG cells. Inhibition of 2-HGproduction by Compound 1 in the IDH1^(R132H) and IDH1^(R132C) U87 linesgave IC₅₀ values of 9.0 and 39.0 nM, respectively, which are in closeagreement with those seen in the HT1080 and HCT-116 (Table 1). Inaddition, Compound 1 potently inhibited 2-HG production in IDH1^(R132G),IDH1^(R132S) and IDH1^(R132L) expressing cells with IC₅₀ values of 5.6,9.2, and 41.7 nM, respectively, suggesting that Compound 1 is a potentinhibitor against a broad spectrum of IDH1^(R132) mutants. In agreementwith the previous cell lines studies, Compound 1 was found to haveminimal effects on the proliferation of mIDH1 expressing U87MG cells at10 μM.

Additional results are shown in Table 10, relative to the IC₅₀ valueobtained for R132H IDH1 mutated enzyme. Referring to data in Table 10,Compound 1 was found to selectively inhibit the enzymatic activity ofthe IDH1 R132H and IDH1 R132C mutations with an IC₅₀ value within afactor of about 5 (i.e., the IC₅₀ value measured for IDH1 R132C mutantenzyme was about 5 times higher than the IC₅₀ measured in the IDH1 R132Hmutated enzyme). The selectivity of Compound 1 against other IDHisozymes was also tested utilizing diaphorase coupled assays employingeither wild-type IDH1 or one of 2 alternate mutated forms of IDH2,namely IDH2 R172K and IDH2 R140Q.

TABLE 10 Relative Enzymatic IC₅₀ Target (Average +/− SEM, nM) IDH1 R132H1.0 (±6.6%) IDH1 R132C 5.1 (±6.1%) Wild Type IDH1     922 IDH2R172K >1,000 IDH2 R140Q >4,000 (no activity measured)

Compound 1 had comparatively very weak activity against wild type IDH1(IC₅₀ value of about 922 times greater than the IC₅₀ value measured forIDH1 R132H). Compound 1 also demonstrated very weak activity againstIDH2 R172K that was more than 1,000 greater than the IC₅₀ value measuredfor IDH1 R132H. Compound 1 did not show any inhibition of IDH2 R140Q upto a concentration of 100 μM. These selectivity data indicate thatCompound 1 is a potent and selective inhibitor of IDH1 R132 mutations.

Example 3 In Vitro Activity of Compound 1 as a R132X mIDH-1 Inhibitor

In in vitro biochemical assays, Compound 1 significantly inhibitedmutated IDH1-R132H and IDH1-R132C proteins. In contrast, Compound 1displayed little or no inhibitory activity in biochemical assays ofwild-type IDH1 protein or various mutated IDH2 proteins found in humancancers. Compound 1 suppressed 2-HG production in naturally occurringand genetically engineered cell lines expressing five different mutatedIDH1 proteins (R132H, R132C, R132G, R132L, and R132S) with IC₅₀ valuesbelow about 0.5 micromolar. In addition, Compound 1 has displayedrelevant levels of activity against multiple clinically relevant,mutated forms of IDH1, of which IDH1-R132H and IDH1-R132C are the mostprevalent in hematologic and solid tumor malignancies. However, Compound1 did not display appreciable activity against wild-type IDH1 or mutatedIDH2.

The cellular potency of Compound 1 in suppressing intracellular 2-HGlevels was determined in cell lines expressing five different mutatedIDH1 proteins found in human cancers (R132H, R132C, R132G, R132L,R132S). The human fibrosarcoma cell line HT-1080 harbors a naturallyoccurring heterozygous IDH1-R132C mutation. The human colorectalcarcinoma cell line HCT 116 is wild type for IDH1, but heterozygousmutations coding for IDH1-R132H or -R132C were introduced by knock-ininto the endogenous IDH1 gene locus. Finally, the human astrocytoma cellline U-87 MG is also wild type for IDH1, but expression of fivedifferent mutated IDH1 proteins was achieved by stable transfection.

The parental HCT116 line (colon) line does not produce high levels of2-HG, but the variants used herein (X-MAN HCT-116 lines obtained fromHorizon Discovery Ltd.) are engineered to knock-in a heterozygousmutation of either IDH1 R132H or IDH1 R132C. This recapitulates thecellular context in mIDH1 cancer cells where there are both wild typeand mutant IDH1 subunits that together form a heterodimer that isresponsible for the production of elevated levels of 2-HG. Thesemodified lines can be used as models of IDH1 mutant disease.

Each of these cell lines was treated with Compound 1 for 24 hr, andintracellular 2-HG levels were determined by mass spectroscopy. Compound1 suppressed 2-HG production in each cell line, with IC₅₀ values rangingfrom <10 to <150 nM. Table 11 below indicates 2-HG IC₅₀ values: below150 nM (“+”), below 100 nM (“++”), below 50 nM (“+++”) and below 10 nM(“++++”).

TABLE 11 Cell Line 2-HG IC50 (nM)* HT-1080(IDH1-R132C/+) ++ HCT116(IDH1-R132H/+) +++ HCT 116(IDH1-R132C/+) + U-87 MG/IDH1-R132H +++U-87 MG/IDH1-R132C ++ U-87 MG/IDH1-R132G ++++ U-87 MG/IDH1-R132L +++U-87 MG/IDH1-R132S ++++ Wild type-IDH1 IC₅₀ (μM) +++ mIDH2-R172K IC₅₀(μM) +++ mIDH2-R140Q IC₅₀ (μM) + *Mean +/− SEM where applicable

Compound 1 is therefore a potent inhibitor of a variety of clinicallyrelevant IDH1 mutations in a cellular context.

In order to optimize the dosing schedule of Compound 1 to achievesustained >90% 2-HG inhibition in mIDH1 in vivo, HCT116-IDH1^(R132H) andHCT116-IDH1^(R132C) xenograft-bearing mice were treated with Compound 1at 25 and 50 mg/kg BID (3 doses). The free drug concentration ofCompound 1 at 12 hour post final dose is above the in vivo IC₉₀ for alldoses, and a greater than 90% reduction of 2-HG levels in tumor wereachieved in each case. The free drug concentration decreased to 3-10×the in vivo IC₅₀ at 24 hour post final dose, and the compound showed80-90% inhibition. There was less than 20 nM free drug concentration intumor at 48 and 72 hours after final dose, and at that point there wasless than 50% 2-HG inhibition in tumor samples, consistent with thereduced level of Compound 1.

In both IDH1 mutated models, the free concentration of Compound 1 wascomparable in plasma and xenograft tumors, and exposures weredose-dependent. In comparison to the vehicle treated group, Compound 1showed a time and dose-dependent inhibition of intratumoral 2-HG levels.At the highest dose tested in these studies (50 mg/kg), Compound 1treatment inhibited 2-HG levels in tumor by >90% for up to 24 hoursafter the last dose in the HCT116-IDH1^(R132H) xenograft model, and tosimilar levels for at least 12 hours in the HCT116-IDH1^(R132C) model.Calculations based upon the percentage of suppression of 2-HGconcentration in tumor versus the free drug concentration in tumor gavein vivo IC₅₀ values of less than 50 nM in both the HCT116-IDH1^(R132H)or HCT116-IDH1^(R132C) models.

Example 4 Comparative Compounds Demonstrated Greater Disparity Between2-HG Inhibition in R132C and R132H IDH-1 Cells, Compared to Compound 1

The comparative activity of each of a series of mIDH-1 inhibitorcompounds including Compound 1 were measured using the cell based assayin Example 3. The ratio of the IC₅₀ values obtained from IDH-1 R132CHCT116 mutant cells (IC₅₀ μM g mean)/IC₅₀ values obtained from IDH-1R132H HCT116 mutant cells (IC₅₀ μM g mean) is provided in Table 4.Compound 1 had the lowest ratio among the tested compounds, indicatingnear equipotent activity of Compound 1 as measured with the R132C andR132H IDH-1 mutant cell assay of Example 3 (using the HCT 116 cellsdescribed in Example 3). Compound 1 showed comparative activityinhibiting 2-HG production from mIDH-1 R132C and R132H cell lines (usingthe assay of Example 3) that was within 5-fold, compared to moredisparate differences in activity ranging from about 8-fold to over 200fold (240) in comparative compounds A-H in Table 12.

TABLE 12 Ratio of IC₅₀ measured for: Compound Structure [IC₅₀ forR132C]/[IC₅₀ for R132H] 1

4.5 A

8.0 B

8.0 C

8.5 D

9.0 E

11.0 F

26 G

30 H

240

Example 5 Determination of Central Nervous System MultiparameterOptimization (CNS MPO)

Central nervous system multiparameter optimization (CNS MPO) may be usedto prioritize compounds based on their likelihood to be brain-penetrant.The scoring function uses six key physicochemical properties (scoringeach parameter on a scale of zero to one) to arrive at a composite scoreranging from 0-6. Higher CNS MPO scores are correlated with a higherlikelihood of a compound being brain-penetrant. The reported CNS MPOscores were calculated following the method reported in: Wager, T. T.,Hou, X., Verhoest, P. R., and Villalobos, A. (2010) Moving beyond rules:The development of a central nervous system multiparameter optimization(CNS MPO) approach to enable alignment of druglike properties. ACS Chem.Neurosci. 1, 435-449.

A summary of the MPO scores for several IDHm inhibitors can be found inTable 13:

TABLE 13 IDH1 IDH1 CNS R132H R132C MPO Predicted Cpd ID Structure IC₅₀(μM) IC₅₀ (μM) Score^(a) BBB+^(b) BBB+^(c) AG-120¹

0.012 0.013 3.69 No No AG-881²

>0.022 >0.022 3.97 Yes Yes IDH305³

0.027 0.028 4.15 Yes Yes IDH889⁴

0.020 0.072 4.50 Yes Yes GSK321⁵

0.0048 0.0038 2.89 nd No Bay1436032⁶

0.015 0.015 3.05 nd No Compound 1^(d)

+++ (0.0212) +++ (0.1138) 5.22 Yes Yes I-1

+++ nd 5.30 nd Yes I-2

++ + 5.35 nd Yes I-3

++ + 5.42 nd Yes I-5

++ + 5.29 nd Yes I-6

++ + 5.29 nd Yes I-11

++ + 5.27 nd Yes I-20

+++ ++++ 4.93 nd Yes I-22

+++ ++++ 4.93 nd Yes I-23

+++ ++++ 3.97 nd Yes I-25^(e)

+++ nd 3.97 nd Yes I-26

++++ ++++ 3.75 nd No I-27

++++ ++++ 4.38 nd Yes I-29

++++ ++++ 4.46 nd Yes For Compound 1 and Compounds I-1 through I-29,WO/2016044789 defines IC₅₀ values for IDH1 R132H as ″++++″: <0.01 μM;″+++″: between 0.01 μM and 0.1 μM; ″++″: from 0.1 μM to 1 μM; and IC₅₀values for IDH1 R132C as ″++++″: <0.1 μM; ″+++″: between 0.1 μM and 1μM; ″++″: from 1 μM to 10 μM; and ″+″: >10 μM ^(a) CNS MPO scorecalculated based on the method described in T. Wager, ACS Chem.Neurosci. (2010), 1, 435-449. ^(b) Literature reported data ^(c)Predicted BBB+: CNS MPO score >3.8 ^(d) WO/2016044789 also reportsCompound 1 (as I-13) as having activity in HCT116 mutant IDH1 R132H andR132C cells as +++ and +++, respectively. ^(e) WO/2016044789 alsoreports I-25 as having activity in HCT116 mutant IDH1 R132H and R132Ccells as ++++ and ++++, respectively. ¹ Popovici-Muller, J., et al.Discovery of AG-120 (Ivosidenib): A First-in-Class Mutant IDH1 Inhibitorfor the Treatment of IDH1 Mutant Cancers. ACS Med. Chem. Lett., 2018,9(4), 300-305. ² Yen, K., et al. Abstract B126: AG-881, a brainpenetrant, potent, pan-mutant IDH (mIDH) inhibitor for use in mIDH solidand hematologic malignancies, AACR-NCI-EORTC International Conference:Molecular Targets and Cancer Therapeutics; Oct. 26-30, 2017;Philadelphia, PA. ³ Cho, Y. S. , et al. Discovery and evaluation ofclinical candidate IDH305, a brain penetrant mutant IDH1 Inhibitor. ACSMed. Chem. Lett. 2017, 8, 1116-1121. ⁴ Levell, J. R., et al.Optimization of 3-pyrimidin-4-yl-oxazolidin-2-ones as allosteric andmutant specific inhibitors of IDH1. ACS Med. Chem. Lett. 2017, 8,151-156. ⁵ Okoye-Okafor, U. C., et al. New IDH1 mutant inhibitors fortreatment of acute myeloid leukemia. Nat. Chem. Biol. 2015, 11, 878-886.⁶ Pusch, S., et al. Pan-mutant IDH1 inhibitor BAY 1436032 for effectivetreatment of IDH1 mutant astrocytoma in vivo. Acta Neuropathologica2017, 133(4), 629-644.

Example 6 Determination of Predicted C_(brain) Ratio of IDHm Inhibitors

The distribution of Compound 1, AG-120 or AG-881 into the brain wasmeasured ex-vivo, individually, in Sprague Dawley rats (n=4/molecule)following a 6 hour intravenous infusion of a 7.5 mg/kg dose. Braintissue and plasma samples were collected at the end of the infusion timeand were processed for bioanalysis via tandem HPLC-mass spectrometryanalysis (LCMS) to determine the total amount of compound present. Inparallel, brain and plasma samples were subjected to equilibriumdialysis as described by N. J. Waters et. al (J. Pharm Sci (2008)97(10):4586-95) to determine the unbound fraction (Table x). The braindistribution or partitioning coefficient (Kpuu) was then calculated asthe ratio of the unbound concentration of drug in brain (Fu, brain) tothe unbound concentration of drug in plasma (Fu, plasma). Similarly, allcalculations of effective plasma concentrations in rodents or humanswere conducted using the unbound/free fraction measured for eachmolecule in each species using conventional equilibrium dialysis asdescribed by Waters.

To determine the projected what the effective free concentration inhumans, the plasma concentrations in tumor bearing mice were measuredusing 90% inhibition in tumors (IC₉₀) of the biomarker 2-HG, as theminimal effective concentration to provide therapeutic benefit. In tumorbearing mice studies the unbound plasma concentration at IC₉₀ wasdetermined to be 90.7 ng/mL (Table 14). In human clinical trials a doseof 150 mg BID for Compound 1 showed a mean plasma concentration(C_(free, avg)) of 171 ng/mL, which when corrected by the expected brainpartitioning (Kpuu=0.42) and free fraction in brain (Fu, brain=0.05)provides an estimate of 3.6 ng/mL unbound concentration in brain. Thetarget effective concentration in human brain based on mouse models (toachieve IC₉₀) is 1.91 ng/mL. Hence Compound 1 partitions into the brainby 2-fold greater than projected levels required to achieve therapeuticbenefit.

TABLE 14 Parameters Compound (ng/mL) 1 AG-120 AG-881 Brain Distribution0.4 0.01 0.5 (Kp, uu rat) *Predicted C_(eff,brain,fu) 1.9 0.7 0.5Predicted Clinical 150 mg 500 mg 50 mg 10 mg C_(Avg)_Brain,fu bid: qd**:qd: qd: 4 0.4 0.03 0.01 Predicted C_(brain) Ratio 2.0 0.6 0.06 0.02(C_(avg)_Brain,fu/ C_(eff)_Brain,fu) *Calculated brain distribution ofCeff,fu plasma in mice **Based Cycle 1 PK, 2-fold decrease over time

Example 7 Testing Compound 1 in Mouse Xenograft Models Using HCT 116Cells with R132C and R132H Mutations

In order to assess the in vivo activity of Compound 1, PK-PD experimentsin mice bearing HCT-116 xenografts (derived from Horizon Discoveryisogenic cell lines harboring IDH1-R132H and IDH1-R132C knock-inmutations) were used to determine the degree of exposure required tosuppress 2-HG levels. Compound 1 was administered toHCT116-IDH1-R132H/+xenograft bearing female BALB/c Nude mice at threeoral doses (12.5, 25, and 50 mg/kg) in 12-hour intervals. Plasma andxenograft tumor samples were collected at 4, 12, and 24 hours post lastdose to determine the exposure of Compound 1 in plasma and tumor, aswell as to measure the inhibition of IDH1 mutant activity in tumor basedon the reduction in levels of 2-HG. In IDH1-R132H/+ xenograft models,the free concentration of Compound 1 was comparable in plasma andxenograft tumors, and exposures were dose-dependent. In comparison tothe vehicle treated group, Compound 1 showed a time and dose-dependentinhibition of 2-HG levels in plasma and in tumor) At the highest dosetested in these studies (50 mg/kg), treatment with Compound 1 inhibited2-HG levels in the tumor by >90% for up to 24 hours after the last dosein the HCT116-IDH1-R132H/+ xenograft model (FIG. 18C), and to similarlevels for at least 12 hours in the HCT116-IDH1-R132C/+ model.Calculations based upon the percentage of suppression of 2-HGconcentration in tumor versus the free drug concentration in tumor gavein vivo IC₅₀ values of 26 nM and 36 nM in the HCT116-IDH1-R132H orHCT116-IDH1-R132C models, respectively. When corrected for unboundlevels of Compound 1, there is an excellent correlation in potency amongthe biochemical assay, cellular assay, and in vivo studies.

TABLE 15 mIDH1-R132H mIDH1-R132C Enzyme Cell 2-HG In vivo 2-HG EnzymeCell 2-HG In vivo 2-HG (nM) (nM) (nM) (nM) (nM) (nM) 17 37 26 100 66 36

In order to optimize the dosing schedule of Compound 1 to achievesustained >90% 2-HG inhibition in mIDH1 in vivo, HCT116-IDH1 R132H andHCT116-IDH1 R132C xenograft-bearing mice were treated with Compound 1 at25 and 50 mg/kg BID (3 doses). The free drug concentration of Compound 1at 12 hour post final dose is above the in vivo IC₉₀ for all doses, anda greater than 90% reduction of 2-HG levels in tumor were achieved ineach case. The free drug concentration decreased to 3-10× the in vivoIC₅₀ at 24 hour post final dose, and Compound 1 showed 80-90% (orgreater) inhibition. There was less than 20 nM free drug concentrationin tumor at 48 and 72 hours after final dose, and at that point therewas less than 50% 2-HG inhibition in tumor samples, consistent with thereduced level of Compound 1.

Briefly, 5×10⁶ HCT-116 IDH1-R132H/+ cells (Horizon Discovery) in PBS wasinoculated subcutaneously at the right flank of the 6 weeks old femaleBALB/c nude mice. When the tumor size reached 360-400 mm³, mice wererandomized by tumor volume into nine mice per group. The tumor bearingmice were treated with vehicle (9:1 PEG400:Ethanol) or Compound 1 forthree doses with 12 hr dosing interval. The dosing volume was 10 μL/g.The plasma samples and tumor samples were collected at 4, 12 and 24 hrpost final dose (N=3 mice per time point) for the subsequent measurementof compound level in plasma and tumor samples and of 2-HG level in thetumor samples by UPLC-MS-MS system.

In a separate dosing example, 5×10⁶ HCT-116 IDH1-R132C/+ cells (HorizonDiscovery) in PBS was inoculated subcutaneously at the right flank ofthe 6-8 weeks old female BALB/c nude mice. When the tumor size reached˜250 mm³, mice were randomized by tumor volume into nine mice per group.The tumor bearing mice were treated with vehicle (9:1 PEG400:Ethanol) orCompound 1 for six doses with 12 hr dosing interval. The dosing volumewas 10 μL/g. The plasma samples and tumor samples were collected at 4, 8and 12 hr post final dose (N=4 mice per time point) for the subsequentmeasurement of compound level in plasma and tumor samples and of 2-HGlevel in the tumor samples by UPLC-MS-MS system.

For each assay, the total concentration of Compound 1 was determined inμM and free Compound 1 concentration was calculated by multiplying thetotal Compound 1 concentration by 0.043 given that Compound 1 is 95.7%protein binding in mouse plasma. The percentage of 2-HG inhibition inindividual tumor sample in the treated groups was normalized to theaverage of 2-HG concentration in the vehicle group at the correspondingsampling time using the following calculation: % 2-HGinhibition=100*(A−B)/A, where A is the average of 2-HG concentration atthe corresponding sampling time, B is the 2-HG concentration in thetumor treated with given dose of Compound 1 and sacked at the givensampling time. The in vivo potency of Compound 1 for suppressing 2-HG intumor is calculated by plotting the percentage of 2-HG inhibitionagainst corresponding free Compound 1 concentration in tumor and fittingthe data with four-parameter logistic equation.

IDH1-R132H Mutation

IDH1-R132H mutation resulted in elevation of 2-HG level in hematologicaland solid cancers. HCT-116 IDH1-R132H/+ xenograft tumor was used toassess the in vivo potency of Compound 1 to suppress 2-HG in tumorlysates. The tumor bearing mice were randomized by tumor size intotwelve mice per group. The mice were treated with Compound 1 at 6.25,12.5, 25, or 50 mg/kg for six doses with dose interval of 12 hr. Theplasma and tumor samples were collected at 4, 8, and 12 hr post lastdose with four mice per time point. The Compound 1 concentration inplasma and tumor samples was analyzed by LC-MS method. The 2-HG level intumor samples was analyzed by LC-MS method. The percentage of 2-HGsuppression in tumor lysate at given dose of Compound 1 was thennormalized to 2-HG level in the vehicle control group. The dose and timedependent 2-HG inhibition by Compound 1 was observed in this study. Thedegree of 2-HG inhibition in tumor lysates was correlated with the freedrug concentration in the corresponding tumor lysate. The calculated invivo potency of Compound 1 to suppress 2-HG in tumor was 26.0 nM.

Upon correcting for unbound Compound 1 concentration, there was a goodcorrelation between the enzymatic, cellular 2-HG, and in vivo 2-HGpotencies of Compound 1 for IDH1-R132H mutant.

IDH1-R132C Mutation

IDH1-R132C mutation resulted in elevation of 2-HG level in hematologicaland solid cancers. HCT-116 IDH1-R132C/+ xenograft tumor was used toassess the in vivo potency of Compound 1 to suppress 2-HG in tumorlysates. The tumor bearing mice were randomized by tumor size into ninemice per group. The mice were treated with Compound 1 at 12.5, 25, or 50mg/kg for three doses with dose interval of 12 hr. The plasma and tumorsamples were collected at 4, 12, and 24 hr post last dose with threemice per time point. The Compound 1 concentration in plasma and tumorsamples was analyzed by LC-MS method. The 2-HG level in tumor sampleswas analyzed by LC-MS method. The percentage of 2-HG suppression intumor lysate at given dose of Compound 1 was then normalized to 2-HGlevel in the vehicle control group. The dose and time dependent 2-HGinhibition by Compound 1 was observed in this study. The degree of 2-HGinhibition in tumor lysates was correlated with the free drugconcentration in the corresponding tumor lysate. The calculated in vivopotency of Compound 1 to suppress 2-HG in tumor was 36.0 nM.

Upon correcting for unbound Compound 1 concentration, there was a goodcorrelation between the enzymatic, cellular 2-HG, and in vivo 2-HGpotencies of Compound 1 for IDH1-R132C mutant.

Results

Given the role of 2-HG in suppressing normal differentiation of mt-IDH1cells (Figueria et al., 2010; Saha et al., 2014), it is hypothesizedthat in order to reverse and maintain this effect, it is necessary toachieve a very high degree of target inhibition on a continuous basis.Therefore, in order to optimize the dosing schedule of Compound 1, it isnecessary to achieve sustained >90% 2-HG inhibition in mt-IDH1 in vivo.For the HCT116-IDH1^(R132H) xenograft assay, the 12 and 24 hour timepoints were chosen to reflect the compound level and corresponding 2-HGinhibition at the C_(trough) of BID and QD dosing schedules. The 48 and72 hour time points were selected to investigate whether Compound 1 hadlong lasting effects on 2-HG inhibition. The free drug concentration ofCompound 1 at 12 hour post final dose is above the in vivo IC₉₀ for alldoses, and a greater than 90% reduction of 2-HG levels in tumor wereachieved in each case. The free drug concentration decreased to 3-10×the in vivo IC₅₀ at 24 hour post final dose, and the compound showed80-90% inhibition. There was less than 20 nM free drug concentration intumor at 48 and 72 hours after final dose, and at that point there wasless than 50% 2-HG inhibition in tumor samples, consistent with thereduced level of Compound 1. These data support the premise thatconstant target coverage by a significant margin is required to achievesustained 2-HG inhibition. This experiment also suggests that a BIDschedule is the preferred dosing regimen for Compound 1 in order tocontinuously achieve >90% 2-HG inhibition. This level of inhibition hasrecently been correlated to clinical efficacy with AG-221 in mt-IDH2harboring AML patients (Fan et al., 2014)).

The present disclosure contemplates, among other things, recognitionthat the total concentration (C_(eff)) of Compound 1 must be above 1652ng/mL in human patients in order to achieve 90% inhibition of 2-HG andabove 2000 ng/mL to achieve greater than 90% inhibition of 2-HG. C_(eff)was determined using assays outlined in Example 7. In two separate mouseexperiments, HCT-116 IDH1-R132H/+ xenografts and HCT-116 IDH1-R132C/+xenograft tumor were used to assess the in vivo potency of Compound 1 tosuppress 2-HG in tumor lysates. Compound 1 concentration in plasma andtumor samples and 2-HG level in tumor samples was measured. The degreeof 2-HG inhibition in tumor lysates was correlated with the free drugconcentration in the corresponding tumor lysate (see FIG. 18D). Giventhe role of 2-HG in suppressing normal differentiation of mt-IDH1 cells(Figueria et al., 2010; Saha et al., 2014), the present disclosurehypothesized that in order to reverse and maintain this effect, it isnecessary to achieve a very high degree of target inhibition withCompound 1 on a continuous basis. It was previously proposed that >90%inhibition of 2-HG correlates to clinical efficacy in mt-IDH2 harboringAML patients (FAN, B. et al., Evaluation of thepharmacokinetic/pharmocodynamic (PK/PD) relationships of an oral,selective, first-in-class, potent IDH1 inhibitor, AG-221, from a phase 1trial in patients with advanced IDH2 mutant positive hematologicmalignancies, Blood, 124: 3737, 6 pages (2014)). Using the curve fromFIG. 18D, the level of free drug concentration of Compound 1 wasdetermined to be 0.256 μM in order to achieve 90% inhibition of 2-HG.

Using a rapid equilibrium dialysis approach, the plasma protein bindingfor a human patient was determined to be 94.5%. (Waters, N. J., et al.(2008)). Validation of a rapid equilibrium dialysis approach for themeasurement of plasma protein binding. J Pharm Sci 97(10): 4586-95.)Accordingly, the total concentration (C_(eff)) can be determined:0.256/((100−94.5)/100) =4.65 μM=1652 ng/mL.

Example 8 Compound 1 Penetrates the Blood Brain Barrier in Murine Models

The blood brain barrier penetration and free brain exposure of Compound1 was investigated in the male CD-1 mouse (FIG. 5). The brain exposureof Compound 1 following oral dosing (5 mg/kg) was evaluated. Following a5 mg/kg oral dose in CD1 mice, the brain to plasma ratio of Compound 1was found to be 0.24 suggesting that the compound has reasonable brainpenetration characteristics, and that at higher doses Compound 1 wouldhave the potential to achieve therapeutic brain levels.

This was confirmed by dosing at 100 mg/kg (FIG. 6), where the brain toplasma ratio was determined to be 0.38. Importantly, following a single100 mg/kg PO dose, the free Cmax concentration of Compound 1 was about200 nM (approximately ten fold IC₅₀), dropping to about 130 nM (aboutsix fold IC₅₀) at the 7 hour time point, which is consistent withexposures that produced >90% suppression of 2-HG in both HCT116 (IDH-1R132H) and HCT116 (IDH-1 R132C) xenograft PK-PD studies. Based onassessment in the mouse, Compound 1 crosses the blood-brain barrier toreach free concentration levels in the brain consistent withpharmacological activity. An oral 100 mg/kg dose gives a free Cmax inbrain of 200 nM, dropping to about 130 nM at the 7 hr time point.

Example 9 Testing Compound 1 in Cynomolgus Monkey Models

Animal Acquisition and Acclimation

A total of 22 male and 22 female experimentally naïve cynomolgusmonkeys, approximately 2 years and 7 months to 3 years and 11 months ofage at transfer, were transferred from the stock colony. The animalswere originally received from Worldwide Primates Inc.

Animals were quarantined upon arrival and quarantine activities,including intrapalpebral tuberculin skin tests, were performed. Theanimals were considered suitable prior to being released fromquarantine. During acclimation as part of the stock colony, the monkeyswere examined by a staff veterinarian, weighed, and observed daily withrespect to general health and any signs of disease.

Randomization, Assignment to Study, and Maintenance

Using a standard, by weight, measured value randomization procedure, 20male and 20 female animals (weighing 2.50 to 3.15 kg and 2.35 to 3.20kg, respectively, at randomization) were assigned to the control and 3treatment groups. Animals assigned to study had body weights within ±20%of the mean body weight for each sex. Extra animals obtained for thestudy, but not placed on study, were transferred to the stock colony.Each animal was assigned an animal number to be used in the datacollection system and was implanted with a microchip bearing a uniqueidentification number. Each animal was also identified by a permanenttattoo with the vendor animal number. The individual animal number,implant number, tattoo number and study number comprised a uniqueidentification for each animal. Each cage was identified by the animalnumber, study number, group number, and sex.

Administration

Male and female cynomolgus monkeys were assigned to four groups. Animalswere assigned to the study as indicated below in Table A. Six animalsper sex in Group 1 were administered vehicle control article only. Fouranimals per sex in Groups 2 and 3, and 6 animals per sex in Group 4 wereadministered test article. Animals in Groups 2 through 4 were dosed for28 days. 2 animals/sex in Groups 1 & 4 were assigned for 28-day recoveryassessment. Animals were dosed via oral gavage twice daily, every 12hours, at a volume of 10 mL/kg/dose (20 mL/kg/day). Animals in Groupstwo through four were administered 30, 100/50, or 300/200/100 mg/kg/day(15, 50/25, or 150/100/50 mg/kg/dose) respectively. The dose levels werelowered for Group 3 and Group 4 animals based clinical observationsduring the dosing phase. The vehicle control article was KolliphorEL:Tween 80 (70:30, v/v). Animals designated for recovery sacrifice (2animals/sex in Groups 1&4) underwent 4 weeks of recovery assessment.

TABLE A Group Assignments Dose Dose Dose Group Level Level Volume DoseNumber of Num- (mg/kg/ (mg/kg/ (mL/kg Concentration Animals ber day)dose) dose) (mg/mL) Male Female 1 0^(a) 0 10 0 6^(b) 6b 2 30 15 10 1.5 44 3 100/50^(c) 50^(c) 10 5 4 4 4 300/200/ 150/100^(d) 10 15/10^(d) 6^(b)6^(b) 100^(d) ^(a)Animals in Group 1 received the vehicle, Kolliphor EL:Tween 80 (70:30, v/v). ^(b)Two animals/sex were maintained for a 28-dayrecovery period. ^(c)Beginning on Day 14, the 50 mg/kg/dose twice daily(BID) dose (100 mg/kg/day) was reduced to 50 mg/kg/dose once daily (50mg/kg/day) for the remainder of the study. ^(d)Animals at 300 mg/kg/daywere placed on a dosing holiday beginning with the second dose on Days 4through Day 11. The 150 mg/kg/dose twice daily (BID) dose (300mg/kg/day) was reduced to 100 mg/kg/dose twice daily (200 mg/kg/day) ata concentration of 10 mg/mL for Days 12 and 13. Beginning on Day 14, the100 mg/kg/dose twice daily (BID) dose (200 mg/kg/day) was reduced to 100mg/kg/dose once daily (100 mg/kg/day) for the remainder of the study.

Assessment of toxicity was based on mortality, clinical observations,body weights, food consumption, ophthalmic observations,electrocardiographic (ECG) measurements, and clinical and anatomicpathology. Blood samples were collected for toxicokinetic evaluations.

Electrocardiographic Assessment

With the exception of one animal in the high dose group, there was noeffect of Compound 1 on qualitative ECG parameters. Frequent ventricularpremature complexes (Day 1, 3-4 hours post-dose) and ventriculartachycardia (Day 28 pre-dose and 3-4 hours post-dose) were observed inone animal following administration of the 300/200/100 mg/kg/day dose.As these ventricular arrhythmias are not considered normal variants andwere observed following the high dose, these findings may have been testarticle-related. Noteworthy effects on quantitative ECG parameters werelimited to QTc interval duration. When evaluated statistically by sex,mean QTc interval duration was longer (vs. concurrent vehicle controlvalue) in 300/200/100 mg/kg/day males at the Day 1 post-dose intervaland at the Day 28 pre-dose and post-dose intervals and in 100/50mg/kg/day females at the pre-test and Day 28 intervals. The differencein 100/50 mg/kg/day females was not considered to be testarticle-related as it was present prior to initiation of test articleadministration. In 300/200/100 mg/kg/day males, the increase in mean QTcinterval duration may have been test article-related as it was observedat the highest dose level and exhibited a progressive increase withcontinued dosing. The magnitude of change from pretest values was mildto moderate (Day 1 post-dose: 7.14%; Day 28 pre-dose: 7.73%; Day 28post-dose: 10.6%). Compared to pretest values, the magnitude of theincrease in QTc interval duration in males at the Day 28 post-doseinterval was 10.61%, which approximates the 10% change seen in theJapanese QT PRODACT studies (Ando, K., Hombo, T., Kanno, A., Ikeda, H.,et al. QT PRODACT: in vivo QT assay with a conscious monkey forassessment of the potential for drug induced QT interval prolongation. JPharmacol Sci. 2005; 99(5):487-500) of drugs known to cause QTprolongation in people. The effect on QTc interval duration wasreversible, not being present at the recovery interval. The thresholdC_(max) plasma concentration in a monkey that experienced an averagegroup QT_(c) prolongation of 10.6% was 7840 ng/mL.

Conclusion

Cynomolgus monkeys tolerated oral doses of Compound 1 twice daily at 15mg/kg/dose (30 mg/kg/day) and once daily at 50 mg/kg/day. Compound1-related post-mortem findings at the end of the treatment periodincluded macroscopic findings of black discoloration in the liver of 3/4males and 2/4 females at 300/200/100 mg/kg/day, which correlated tomultinucleated cells in the sinusoids. Lower thymus weights, whichcorrelated to an increased incidence and/or severity of lymphoiddepletion, were observed in Compound 1-treated animals and wereconsidered secondary to stress and ill-health. Microscopically, Compound1 was associated with multinucleated cells in the sinusoids of the liverand mucosal atrophy of the intestinal tract in males and females at100/50 and/or 300/200/100 mg/kg/day. The histological changes in theliver were considered adverse and correlated with a number of the serumchemistry changes. At the recovery interval, intestinal changes wereabsent, suggesting reversibility, and multinucleated cells in thesinusoids of the liver were decreased in severity, indicating partialrecovery. Test article-related increase in the mean QT_(c) level wasobserved at the highest dose level 300/200/100 mg/kg/day in the malemonkeys. The threshold plasma C_(max) concentration in a monkey thatexperienced an average group QT_(c) prolongation of 10.6% was 7840ng/mL.

The no-observed-adverse effect-level (NOAEL) for Compound 1 wasconsidered to be 30 mg/kg/day (15 mg/kg/dose BID). Systemic exposure(Cmax and AUCTIast; combined-sex) at the NOAEL on Day 28 was 2490 ng/mLand 23600 ng·h/mL, respectively. Based on the expected reversibility ofadverse hepatic findings, 50 mg/kg/day was considered the highestnonseverely toxic dose (HNSTD). Systemic exposure (Cmax and AUCTIast;combined-sex) at the HNSTD on Day 28 was 4350 ng/mL and 53900 ng·h/mL,respectively.

Example 10 A Phase 1 Dose Escalation Study of the mIDH-1 Inhibitor,Compound 1, in Patients with AML or Myelodysplastic Syndrome MDS

Isocitrate dehydrogenase 1 mutations (m/DH-1) occur in 7-14% of AMLpatients (“pts.”) and 3% of MDS pts. Compound 1 is a highly potent,selective small molecule inhibitor of mIDH-1 without anticipated CYP orQTc liabilities at the recommended phase 2 dose. Compound 1 was testedin a Phase 1/2 study to evaluate the safety, efficacy, PK, and PD ofCompound 1 as a single agent or in combination with azacitidine orcytarabine.

FIG. 13A illustrates the summary of cohorts from a phase 1 study inIDH1m AML and MDS, described in this example. The Phase 1/2 study ofCompound 1 was initiated to evaluate the safety, PK/PD, and clinicalactivity of Compound 1 alone or in combination with azacitidine (AZA) orcytarabine in mIDH-1 AML/MDS pts. In the phase 1 portion of the study,Compound 1 was dose escalated in a 3+3 design to define the maximumtolerated doses (MTDs) or maximum evaluated doses (MEDS) as asingle-agent (SA) and in combination with azacitidine (CO) followed byexpansion cohorts. Doses evaluated were 150 mg QD (SA, CO), 300 mg QD(SA), and 150 mg BID (SA, CO). Safety was assessed by incidence andseverity of treatment emergent AEs (TEAEs) for all pts and efficacyderived by IWG criteria (2003 AML and 2006 MDS) based on investigatorassessment for evaluable pts.

FIG. 15 illustrates cycle 1, grade 2 IDH-DS is resolved withdexamethasone and hydroxyurea. C2D1: CRp (1% blasts) to SCT in MLFS,during the clinical trial of Example 10.

Subject Inclusion Criteria Can Include:

-   -   1. Pathologically proven AML or intermediate, high-risk, or very        high risk MDS as defined by the World Health Organization (WHO)        criteria or Revised International Prognostic Scoring System        (IPSS-R) which is relapsed or refractory (R/R) to standard        therapy and/or for which standard therapy is contraindicated or        which has not adequately responded to standard therapy, e.g.        Compound 1 as a single agent: patient has received no prior IDH1        inhibitor or Compound 1 in combination with Aza: patients are        treat-naïve & eligible for Aza, patients can have received IDH1m        inhibitor or HMA.    -   2. Patients must have documented IDH1-R132 gene-mutated disease        as evaluated by the site.    -   3. Good performance status.    -   4. Good kidney and liver function.    -   5. Baseline QTcF 450 msec.    -   6. No exclusions for concomitant medications.        Exclusion Criteria:

-   1. Patients with symptomatic central nervous system (CNS) metastases    or other tumor location (such as spinal cord compression, other    compressive mass, uncontrolled painful lesion, bone fracture, etc.)    necessitating an urgent therapeutic intervention, palliative care,    surgery or radiation therapy.

-   2. Congestive heart failure (New York Heart Association Class III    or IV) or unstable angina pectoris. Previous history of myocardial    infarction within 1 year prior to study entry, uncontrolled    hypertension or uncontrolled arrhythmias.

-   3. Pulmonary disease (e.g. COPD, asthma, etc) that is not controlled    (moderate to severe symptoms) with current medication.

-   4. Active, uncontrolled bacterial, viral, or fungal infections,    requiring systemic therapy.    Treatment/Intervention Plan

Compound 1 was administered as a single agent or in combination withazacitidine or cytarabine. Compound 1 was supplied as 50 mg or 150 mgcapsules and was administered per the protocol defined frequency anddose level. Azacitidine was administered per site's standard of care.Cytarabine will be administered per site's standard of care.

The Phase 1 stage of the study was split into 2 distinct parts: a doseescalation part, which will utilize an open-label design of Compound 1(single agent), or Compound 1+azacitidine (combination agent), orCompound 1+cytarabine (combination agent) administered via one or moreintermittent dosing schedules followed by a dose expansion part. Thedose expansion part will enroll patients in up to 5 expansion cohorts,exploring single-agent Compound 1 activity as well as combinationactivity with azacitidine or cytarabine. Patients may receive only asingle dose of study drug (single-agent arm and combination arm) onCycle 1 Day 1. Following the completion of the relevant Phase 1 cohorts,Phase 2 begins enrollment. Patients are enrolled across 6 differentcohorts, examining the effect of Compound 1 (as a single agent) andCompound 1 with azacitidine (combination) on various AML/MDS diseasestates. Conditions examined include acute myeloid leukemia (also knownas acute myelogenous leukemia) and myelodysplastic syndrome.

TABLE 16 Arms and Interventions of Phase 1 Trial. Arms AssignedInterventions Experimental: PH1 Dose Escalation & Drug: Compound 1Expansion Compound 1 Compound 1 is supplied as 50 mg or 150 mg capsulesand is administered per the protocol defined frequency and dose levelExperimental: PH1 Esc. and Drug: Compound 1 Exp. Compound 1 +Azacitidine Compound 1 is supplied as 50 mg or 150 mg capsules and isadministered per the protocol defined frequency and dose level Drug:Azacitidine (Vidaza) azacitidine is administered per site's standard ofcare Experimental: PH1 Esc. and Drug: Compound 1 Exp. Compound 1 +Cytarabine Compound 1 is supplied as 50 mg or 150 mg capsules and isadministered per the protocol defined frequency and dose level Drug:Cytarabine low-dose cytarabine are administered per site's standard ofcare Experimental: PH2 Cohort 1 Compound 1 Drug: Compound 1 Single AgentRelapsed or Compound 1 is supplied as 50 mg or 150 mg Refractory (R/R)AML capsules and is administered per the protocol defined frequency anddose level Experimental: PH2 Cohort 2 Compound 1 Drug: Compound 1 SingleAgent Compound 1 is supplied as 50 mg or 150 mg AML/MDS in morphologiccomplete capsules and is administered per the remission or completeremission with protocol defined frequency and dose level incompleteblood count recovery (CR/CRi) after cytotoxic-containing inductiontherapy with residual IDH-R132 mutation Experimental: PH2 Cohort 3Compound 1 Drug: Compound 1 Single Agent Compound 1 is supplied as 50 mgor 150 mg R/R AML/MDS, previously treated with an capsules and isadministered per the IDH1 inhibitor protocol defined frequency and doselevel Experimental: PH2 Cohort 4 Drug: Compound 1 Compound 1 +Azacitidine Compound 1 is supplied as 50 mg or 150 mg R/R AML/MDS thatare naïve to prior capsules and is administered per the hypomethylatingtherapy and IDH1 inhibitor protocol defined frequency and dose leveltherapy Drug: Azacitidine (Vidaza) Azacitidine is administered persite's standard of care Experimental: PH2 Cohort 5 Drug: Compound 1Compound 1 + Azacitidine Compound 1 is supplied as 50 mg or 150 mg R/RAML/MDS that have inadequately capsules and is administered per theresponded or have progressed immediately protocol defined frequency anddose level preceeding hypomethylating therapy Drug: Azacitidine (Vidaza)Azacitidine is administered per site's standard of care Experimental:PH2 Cohort 6 Drug: Compound 1 Compound 1 + Azacitidine Compound 1 issupplied as 50 mg or 150 mg R/R AML/MDS that have been previouslycapsules and is administered per the treated with single-agent IDH1inhibitor protocol defined frequency and dose level therapy as theirlast therapy prior to study Drug: Azacitidine (Vidaza) enrollmentazacitidine is administered per site's standard of care

Following the completion of Phase 1, Phase 2 enrollment began. Patientswere enrolled across 6 different cohorts, examining the effect ofCompound 1 (as a single agent) and Compound 1+azacitidine (combination)on various AML/MDS disease states. The Phase 2 cohorts are summarized inTable 17 below:

TABLE 17 Cohort Patient Population Intervention I Patients with relapsedor Recommended phase II dose refractory (R/R) AML (“RP2D”) of Compound 1as a single-agent II Patients with AML/MDS in RP2D of Compound 1 as amorphologic complete single-agent remission or complete remission withincomplete blood count recovery (CR/CRi) after cytotoxic-containinginduction therapy with residual IDH-R132 mutation III Patients with R/RAML/MDS, RP2D of Compound 1 as a previously treated with an single-agentIDH1 inhibitor IV Patients with R/R AML/MDS RP2D of Compound 1 in thatare naïve to prior combination with azacitidine hypomethylating therapyand IDH1 inhibitor therapy V Patients with R/R AML/MDS RP2D of Compound1 in that have inadequately combination with azacitidine responded orhave progressed immediately preceding hypomethylating therapy VIPatients with R/R AML/MDS RP2D of Compound 1 in that have beenpreviously combination with azacitidine treated with single-agent IDH1inhibitor therapy as their last therapy prior to study enrollmentPrimary Outcome Measures

The outcome of the study can be evaluated using the following criteria:

-   1. Maximum Tolerated Doses (MTDs) or Maximum Evaluated Doses (MEDS)    [Phase 1]. Time Frame: Within first 4 weeks of treatment.-   2. Number of Participants with a Dose Limiting Toxicity (DLT) [Phase    1]. Time Frame: Within first 4 weeks of treatment. DLT Criteria can    include:    -   ≥Gr 3 non-hematologic toxicity or laboratory finding    -   Gr 4 hematologic toxicity by Day 42 in absence of disease    -   Inability to tolerate at least 75% of Cycle 1 treatment-   3. Doses recommended for future studies [Phase 1]. Time Frame:    Within first 4 weeks of treatment.-   4. Complete Response (CR, CRi, MLFS, Marrow CR) Rate of Compound 1    as a single-agent or in combination with Azacitidine in patients    with AML/MDS [Phase 2]. Time Frame: As per IWG Response Assessment    Guidelines for AML and MDS based on investigator's assessment    through study completion, e.g. modified IWG AML 2003/MDS 2006.    Secondary Outcome Measures

The outcome of the study can also be evaluated using the followingcriteria:

-   1. Area under the plasma concentration versus time curve (AUC)    [Phase 1 and Phase 2]. Time Frame: Blood samples for PK analysis    collected at multiple visits during the first 60 days of treatment    and on day 1 of all cycles following the first 30 days.-   2. Peak Plasma Concentration (Cmax) [Phase 1 and Phase 2]. Time    Frame: Blood samples for PK analysis collected at multiple visits    during the first 60 days of treatment and on day 1 of all cycles    following the first 30 days.-   3. Time of peak plasma concentration (Tmax) [Phase 1 and Phase 2].    Time Frame: Blood samples for PK analysis collected at multiple    visits during the first 60 days of treatment and on day 1 of all    cycles following the first 30 days.-   4. Time for half of the drug to be absent in blood stream following    dose (T 1/2) [Phase 1 and Phase 2]. Time Frame: Blood samples for PK    analysis collected at multiple visits during the first 60 days of    treatment and on day 1 of all cycles following the first 30 days.-   5. Rate at which drug is removed from blood stream (CL/F) [Phase 1    and Phase 2]. Time Frame: Blood samples for PK analysis collected at    multiple visits during the first 60 days of treatment and on day 1    of all cycles following the first 30 days.-   6. Rate of drug distribution within the blood stream (Vd/F) [Phase 1    and Phase 2]. Time Frame: Blood samples for PK analysis collected at    multiple visits during the first 60 days of treatment and on day 1    of all cycles following the first 30 days.-   7. Reduction of 2-HG levels in plasma [Phase 1 and Phase 2]. Time    Frame: Blood samples for PK/PD analysis collected at multiple visits    during the first 60 days of treatment and on day 1 of all cycles    following the first 30 days.-   8. Evidence of antileukemic or antimyelodysplastic activity of    Compound 1 as determined by complete response (CR), CRi (complete    remission with incomplete hematologic recovery), morphologic    leukemia-free state (MLFS), Marrow CR, partial remission (PR), and    stable disease (SD) as a single-agent or in combination with    azacitidine or cytarabine [Phase 1]. Time Frame: As per IWG Response    Assessment Guidelines for AML and MDS based on investigator's    assessment through study completion.-   9. Incidence and severity of adverse events, clinical laboratory    abnormalities, and changes in ECG parameters as assessed by CTCAE    v4.0 as a single-agent or in combination with azacitidine [Phase 2].    Time Frame: Safety will be assessed from time of first dose through    28 days post last dose.-   10. Additional measures of antileukemic or antimyelodysplastic    activity as determined by CRh, Overall Response (OR), and Stable    Disease of Compound 1 alone or in combination with azacitidine    [Phase 2]. Time Frame: As per IWG Response Assessment Guidelines for    AML and MDS based on investigator's assessment through study    completion.-   11. Time to Response (TTR) [Phase 2]. Time Frame: From first dose of    study drug through time of first response by blood recovery count.-   12. Duration of Response (DOR) [Phase 2]. Time Frame: From time of    first response by blood recovery count through relapse.-   13. Event-Free Survival (EFS) [Phase 2]. Time Frame: From time of    entry on study through progression.-   14. Overall Survival (OS) [Phase 2]. Time Frame: From time of entry    on study through death or date last known alive at end of follow-up.    Disease History and Baseline Characteristics of Exemplary    Participants

A summary the disease history and baseline characteristics of exemplaryparticipants is shown in Table 18.

TABLE 18 Compund 1 + Compund 1 Azacitidine Characteristic n = 31 n = 41Age, median (range), years 71 (35-87) 66 (31-88) Female, % 52 51 ECOGPS-0/1/2, % 25/65/10 32/51/17 IDH1 mutation type R132-, n C (13)/H (9)/S(4)/ C (22)/H (12)/S (4)/ G (4)/L (1) G (2)/Others (1*) AML, n 25 35Relapsed (>12 mo) 14  1 Relapsed (≤12 mo)  4 10 Refractory  8 15Treatment-naïve  3  9 Secondary AML  3 12 t-AML  1  1 Prior regimens,median  2 (0-10)  3 (0-6) (range) Prior HMA/ no prior  6/3/1 IDHminhibitor/Both IDHm inhibitor MDS, n  6  6 Relapsed/Refractory  4  2Treatment naïve  2  4 Prior regimens, median  1 (0-4)  1 (0-4) (range)Prior HMA/ no prior  2/0/0 IDHm inhibitor/Both IDHm inhibitor *Onepatient with a different non-R132 mutation variantResults

In the human clinical trial described in Example 10, when Compound 1 wasadministered as a single agent, it provided a 41% overall response rate(ORR) and 27% CR/CRh in R/R AML. Combination of Compound 1 withazacitidine provided a 46% ORR and 16% CR/CRh in R/R AML.

At the data cutoff, 35 pts with a median of 2 prior regimens (range 1-9)had received Compound 1 in dose-escalation, including 31 single-agent(SA) and 41 azacitidine combination (CO) pts. Steady-state exposure thatexceeded the target IC₉₀ for mIDH-1 was achieved at 150 mg BID,resulting in a reduction of 2-HG to normal levels in the majority ofpts. Furthermore, administration of Compound 1, both as SA and in CO at150 mg BID, enabled all pts to achieve the target Css that exceeded theIC₉₀ for mIDH-1 while staying below the exposures projected from themonkey toxicology studies that could be associated with QT prolongation.PK data were available through Cycle 10; steady state plasma drug levelswere maintained at target Css over the evaluated period. A reduction of2-HG was observed across all dose levels, with pts receiving 150 mg BIDhaving a median within the normal 2-HG limits.

As shown in FIG. 16A, transfusion independence (platelets and/or redblood cells) was observed in all response categories in AML patientstreated with Compound 1 as a single agent who were transfusion-dependentat baseline. As shown in FIG. 16B, significant reduction in bone marrowblast content was observed in patients with a CR/CRi response per IWGresponse criteria. Reductions in bone marrow blast content were alsoobserved in patients in the absence of IWG response (stable disease)including 1 subject (denoted with t in FIG. 16B) with bone marrow blastcount <5% but with blasts present in peripheral blood.

The responses per Investigator assessment per modified IWG aresummarized in Table19:

TABLE 19 Responses to Compound 1 as a single agent. R/R AML All AML &MDS Response N = 22 N = 31 ORR, n (%)* 9 (41) 11 (35) [95% Cl] [21, 64][19, 55] CR, n (%) 4 (18)  5 (16) CRh, n (%) 2 (9)  2 (6) CRi, n (%) 3(14)  4 (13) PR, n (%) 0  0 MLFS, n (%) 0  0 SD, n (%) 5 (23) 11 (35)PD, n (%) 1 (5)  1 (3) NE, n (%) 7 (32)  8 (26)

As shown in FIG. 17A, transfusion independence (platelets and/or redblood cells) was observed in all response categories in AML patientstreated with Compound 1 and azacitidine who were transfusion-dependentat baseline. As shown in FIG. 17B, reduction in bone marrow blastcontent was observed in patients with a CR/CRi response per IWG responsecriteria. Reductions in bone marrow blast content were also observed inpatients in the absence of IWG response (stable disease) including 1subject (denoted with † in FIG. 17B) with bone marrow blast count <5%but with blasts present in peripheral blood. Marrow blast reduction >50%in patients with SD indicates clinical activity in the absence of IWGresponse.

The responses per Investigator assessment per modified IWG aresummarized in Table 20.

TABLE 20 Responses to Compound 1 + azacitidine. R/R AML TN AML All AML &MDS Response N = 26 N = 9 N = 41 ORR, n (%)* 12 (46) 7 (78) 22 (54) [95%Cl] [27, 67] [40, 97] [37, 69] CR, n (%)  3 (12) 2 (22)  8 (20) CRh, n(%)  1 (4) 0  1 (2) CRi, n (%)  5 (19) 4 (44)  9 (22) PR, n (%)  1 (4) 1(11)  2 (5) MLFS, n (%)  2 (8) 0  2 (5) SD, n (%) 11 (42) 1 (11) 14 (34)PD, n (%)  1 (4) 0  1 (2) NE, n (%)  2 (8) 1 (11)  4 (10)Exemplary Patient Disposition

A summary of the patient disposition is shown in Table 21.

TABLE 21 Compound 1 + Compound 1 Azacitidine Characteristic n = 31 (%) n= 27 (%) Patients enrolled, n 31 27 Patients treated 31 (100) 26 (96)Median months on treatment  3 (<1-20 m)  3 (<1-12 m) (range) Patients ontreatment 13 (42) 13 (50) Discontinued treatment 18 (58) 13 (50) Reasonfor Discontinuation Death  6  4 Progressive disease  9  6 Transplant  4 6 Other *  3  3 Investigator decision  2  5 Withdrawal of consent  1  0Adverse event  3  2 * SA: 3 patients discontinued due to resistantdisease/lack of response Combo: 1 patient discontinued for refusal oftreatment, 1 patient to enter hospice, and 1 patient with no reason fordiscontinuation given.Treatment-Emergent Adverse Events (TEAEs) ≥15% All Grades

A summary of TEAEs can be found in Tables 22A and 22B.

TABLE 22A Compound 1, n = 31 (Single Agent) AE Preferred Term All Grade(%) Grade 3/4 (%) Fatigue 13 (42) 1 (3) Nausea 13 (42) 0 Pyrexia 10 (32)2 (7) Pneumonia  7 (23) 5 (16) Vomiting  7 (23) 0 Dyspnea  7 (23) 0Diarrhea  6 (19) 0 Dizziness  6 (19) 1 (3) Decreased Appetite  6 (19) 0Constipation  6 (19) 1 (3) Hypokalemia  6 (19) 2 (7) AST  5 (16) 2 (7)Abdominal  5 (16) 0 distention Epistaxis  5 (16) 1 (3) Headache  5 (16)0 Thrombocytopenia¹  9 (29) 9 (29) Leukocytosis  8 (26) 4 (13) Anemia  7(23) 7 (23) Febrile neutropenia  7 (23) 7 (23) ¹Includes preferred termof platelet count decreased

TABLE 22B Compound 1, n = 41 (Cpd 1 + AZA) AE Preferred Term All Grade(%) Grade 3/4 (%) Nausea 20 (49)  3 (7) Constipation 20 (49)  1 (2)Hypokalemia 15 (37)  4 (10) Fatigue 15 (37)  7 (17) Diarrhea 13 (32)  2(5) Vomiting  9 (22)  0 Headache  9 (22)  1 (2) Decreased Appetite  8(20)  1 (2) Cough  7 (17)  1 (2) Pneumonia  7 (17)  6 (15) Pruritus  7(17)  0 Dysgeusia  7 (17)  0 Creatinine  7 (17)  1 (2) increasedDizziness  6 (15)  1 (2) Asthenia  6 (15)  2 (5) Abdominal Pain  6 (15) 1 (2) Thrombocytopenia¹ 15 (37) 12 (29) Febrile neutropenia 12 (29) 12(29) Neutropenia² 10 (24)  7 (17) Anemia  9 (22)  7 (17) Leukocytosis  9(22)  6 (15) Leukpenia  7 (17)  5 (12) ¹Includes preferred term ofplatelet count decreased ²Includes preferred term of neutrophil countdecreasedAdverse Events (AE) of Interest and Deaths

AEs were assessed per National Cancer Institute's Common TerminologyCriteria for Adverse Events (NCI CTCAE), version 4.03.

No DLTs were observed in dose escalation.

Compound 1 as Single Agent

4 (13%) patients receiving Compound 1 as a single agent exhibitedIDH-DS; all resolved with treatment interruption, dexamethasone,hydroxyurea and supportive care and then resumed treatment with Compound1.

The QTcF maximum change from baseline for patients treated with Compound1 as a single agent is reported in Table 23.

TABLE 23 Maximum Post-Baseline Value ≤480 >480-≤500 >500 >60 msec ΔTreatment Baseline msec msec msec from BL Group Value n (%) n (%) n (%)n (%) Total Com- ≤480 msec 27 (90) 0 0 1 (3.3)^(‡) pound 1 >480-≤500  01 (3.3)^(†) 0 0 SA msec (N = 30) >500 msec  0 0 1 (3.3) 0 Total 27 (90)1 (3.3) 1 (3.3) 1 (3.3)

2 patients with BBB (t in Table 23) enrolled with QTc readings abovenormal at baseline remained stable within the same QTc range ontreatment. One patient (1 in Table 23) had an increase of >60 msec butremained within the normal limits (<450 msec).

9 patients receiving Compound 1 as a single agent died within 30 days oflast dose due to AEs unrelated to treatment with Compound 1. The AEsunrelated to treatment with Compound 1 which resulted in death aresummarized below:

-   -   Progressive Disease (PD) (n=3)    -   Central Nervous System (CNS) Events (n=2)    -   Sepsis (n=1)    -   Cardiac Arrest (n=1)    -   Multiorgan failure (n=1)    -   toxicity to HSCT regime (n=1)        Compound 1 in Combination with Azacitidine

The QTcF maximum change from baseline (BL) for patients treated withCompound 1 and azacitidine in combination is reported in Table 24.

TABLE 24 Maximum Post-Baseline Value ≤480 >480-≤500 >500 >60 msec ΔTreatment Baseline msec msec msec from BL Group Value n (%) n (%) n (%)n (%) Total Com- ≤480 msec 37 (90.0) 2 (5) 2 (5) 4 (10) pound1 + >480-≤500  0 0 0 0 AZA 75 msec mg/m² (N = 41) >500 msec  0 0 0 0Total 37 (90.0) 2 (5) 2 (5) 4 (10)

Referring to Table 24, 4 patients with >60 msec which increased from BLincluded the 2 patients with suspect concomitant medications, 1 patientdeveloped G1 prolongation and 1 patient remained within normal limits.Among the patients with QTcF <480 msec at BL, 2 had values of 480-500msec, and 2 had values >500 msec. 1 of each group (including 1 withpacemaker) had prolonged QTcF before treatment start. The other 2 hadtransient prolongation that normalized once suspect concomitantmedications discontinued 2 AEs of QTcF prolongation reported on study(G2 and G3). These were transient and patients resumed treatment oncesuspect concomitant medications discontinued.

7 patients receiving Compound 1 and azacitidine died within 30 days oflast dose due to AEs unrelated to treatment with Compound 1. The AEsunrelated to treatment with Compound 1 which resulted in death aresummarized below:

-   -   Progressive Disease (PD) (n=3)    -   Central Nervous System (CNS) Events (n=2)    -   Pneumonia (n=1)    -   Gastrointestinal fistula (n=1)

The present disclosure includes, among other things, the novelunderstanding that administration of 300 mg of Compound 1 (e.g., 150 mgBID or 300 mg QD) to a patient or population of patients results in asustained therapeutically effective trough blood plasma concentration(C_(ss)). Such a C_(ss) of Compound 1 resulted in a durable reduction in2-HG plasma level over the course of at least 6 treatment cycles.

As outlined in Example 10, the concentration total plasma concentrationof Compound 1 and the plasma concentration of 2-HG was measured in theblood of patients receiving one of three different dose and doseintervals: 150 mg QD, 300 mg QD or 150 mg BID (either receiving Compound1 as a single agent or in combination with azacitidine as described inthe clinical trial of Example 10, in each category). The 2-HG levelswere measured prior to administration of Compound 1, and then measuredafter administration of Compound 1 up to cycle 2, day 1 after firstreceiving Compound 1 (as the solid form obtained from Example 1).

As shown in FIGS. 19-20, the administration of Compound 1 at 150 mg BIDresulted in a trough blood plasma concentration above 1,652 ng/mL aftercycle 2 of a 28-day treatment cycle as both a single agent and incombination with azacitidine. Additionally, as shown in FIGS. 21-22, areduction of 2-HG level was observed. The decrease in 2-HG concentrationobserved in patients receiving Compound 1 at 150 mg BID or 300 mg QD isunexpectedly better than those who received Compound 1 at 150 mg QD,both in terms of magnitude and variability of effect.

As shown in FIG. 8A-8E, the plasma exposures (steady state blood plasmaconcentration) of Compound 1 were durable (i.e., sustained) throughoutat least a 6 cycle treatment duration. As shown in FIGS. 9A-9C theplasma 2-HG concentrations were reduced within 1 cycle (C2D1) andmaintained throughout the treatment duration.

Example 11 Pharmacokinetics and Pharmacodynamics from Human ClinicalTrial Testing

Prior to Phase 1 trials, evaluation of Compound 1 in in vivo rat and invitro human tissue indicated hepatic metabolism by CYP enzymes (CPY3A4,2C9, 1A1) as the major route of excretion. Animal toxicology studiespredicted the threshold for QTc prolongation risk occurred atexposures >6-fold the in vivo C_(eff) observed in preclinical studiesresulting in 90% reduction in plasma 2-HG.

In human clinical testing described above in Example 10, Compound 1plasma concentrations were determined by a validated and sensitivebioanalytical method. Compound 1 was administered to patients QD (onceper day) and BID (twice per day) as a single agent or in combinationwith azacitidine. On the 1st day of dosing (Cycle 1 Day 1), patientswere administered a single dose of Compound 1 and the plasma wascollected for pharmacokinetics analysis for 24 hours. Based on limitednumbers of evaluable patients (n=5), a plasma half-life of about 60hours was estimated for Compound 1 (steady state by week 2) for 150 mgBID administration of Compound 1, with a Cmax of about 570 ng/mL (wellbelow the levels expected to increase QTcF potential (corrected QTinverval) and an Area Under Curve (0-24 hours) of about 10,000 to 10,050h ng/mL.

A lack of dose proportional response was noted in the Cmax and AUC aftera single dose of Compound 1. Similarly, the steady state trough valuesdid not increase with dose from 150 mg QD to 300 mg QD (see FIG. 10A).The lack of increasing Cmax, and steady trough values is believed to becaused by a dissolution limited absorption, which led to exploration ofdose partition. As demonstrated in FIG. 10A, splitting the 300 mg QDdose to 150 mg BID dose improved the steady state exposure of Compound1, as a single agent and in combination with azacitidine. Notably,concentrations observed in the 150 mg BID dose as a single agent or incombination with azacitidine span the IC₉₀ values for inhibition of allR132X mutations.

Changes in serum 2-HG levels collected at pre-dose and several timepoints through C2D1 (cycle 2, day 1) were measured to assess thepharmacodynamics effect of Compound 1 (See Example 10). A reduction in2-HG was observed across all dosing cohorts with normalization of 2-HGlevels observed with the BID dosing schedule of Compound 1. FIG. 10Bshows the nadir of 2-HG levels in Cycle 1 of Compound 1 dosing observedin patients receiving single agent Compound 1 QD or BID dosing. Themedian level for pre-treatment, QD (150 mg or 300 mg), and BID (150 mg)is about 700, 200, and 70 ng/mL, respectively. Accordingly, the median2-HG levels were reduced by about 10-fold in the 150 mg BID dose ofCompound 1 compared to the 150 mg QD dose of Compound 1.

Example 12 Case Study of Human Clinical Trial Testing of Compound 1

Patient X is 66 y/o female, diagnosed with AML who initially receivedinduction treatment with high dose cytarabine to which the patient wasrefractory. Subsequently, the patient enrolled in a clinical trialstudy, where she was treated with single agent (SA) Compound 1 150 mgBID and achieved a complete remission (CR) after one cycle of treatment(28 days). Patient continued treatment while in CR for 7 additionalcycles. Patient then relapsed and discontinued study treatment.

Patient Y is 62 y/o male, diagnosed with FLT3-positive secondary AML(secondary to MDS). Patient received intensive chemotherapy inductionwith cytarabine and daunorubicin in combination with midostaurin (FLT3inhibitor) but unfortunately was refractory. He enrolled in a clinicaltrial study, where he was treated with Compound 1 150 mg BID incombination with azacitidine for a total of 8 cycles (1 cycle=28 days).He achieved complete remission with IDH1 mutation clearance (CRm) bycycle 6 and discontinued study treatment after cycle 8 to undergo bonemarrow transplant (HSCT).

Patient Z is a 50 year old diagnosed with grade III IDH1m glioma(anaplastic astrocytoma) previously treated with chemotherapy andradiation according to the applicable standard of care. This patient wassubsequently enrolled on the clinical study treated with Compound 1 at150 mg twice daily (BID) each day. Following treatment with Compound 1for 2 cycles (each cycle=28 consecutive days receiving 150 mg Compound 1BID), by MRI, patient was determined by the investigator to haveexperienced a partial response by RANO criteria (≥50% decrease in tumor,no new lesions, on stable dose corticosteroids, no progression ofmeasurable disease). After receiving 2 cycles of Compound 1 (150 mgBID), the patient remains on treatment with 150 mg BID Compound 1 perprotocol.

3 Patients received a Compound 1 at 100 mg once daily (QD) each day.Blood samples were collected every 28 days for measurement of plasmaconcentrations of Compound (single agent) Blood was collected at thefollowing times relative to Compound 1 administration:

-   -   Cycle 1 Day 1: predose, and postdose at 30 minutes, 1, 2, 4, and        8 hours    -   Cycle 1 Days 2, 8, 15, and 22: predose    -   Cycle 2 Day 1: predose, and postdose at 30 minutes, 1, 2, 4, and        8 hours    -   Cycle 2 Day 2: 24 hours after C2D1 dosing (±2 hours) for        patients in dose expansion    -   Cycle 2 Day 4: predose [72 hours after C2D1 dosing (±4 hours)]        for patients in dose expansion only    -   Cycle 2 Day 15: predose    -   Cycle 3 and Beyond: Predose on Day 1 of every cycle

The observed C_(min) associated with this case study can be found inFIG. 19.

Example 13 A Phase 1b/2 Study of Compound 1 in Patients with AdvancedCNS and Solid Tumors with an IDH1 Mutation

Patients having any of the following solid tumors that harbor a IDH1mutation receive Compound 1 (unless otherwise indicated, at a dose of150 mg of the solid form provided in Example 1, administered orally BID)as a single agent or in combination with additional therapies:

-   -   Compound 1 is administered to patients diagnosed with glioma or        chondrosarcoma as a single agent or in combination with        5-azacitidine (also referred to herein as “5-azacytidine” or        “azacytidine” or “azacitidine”);    -   Compound 1 is administered to patients diagnosed with        hepatobiliary cancers as a single agent or in combination with a        PD-1 inhibitor (preferably, nivolumab);    -   Compound 1 is administered to patients diagnosed with        intrahepatic cholangiocarcinoma as a single agent or in        combination with gemcitabine and cisplatin (“GemCis”); and    -   Compound 1 is administered to patients diagnosed with other        non-CNS solid tumors as a single agent.

Each patient has a histologically or cytologically-confirmed IDH1 R132Xgene-mutated advanced solid tumor prior to receiving Compound 1. Inparticular, some patients receiving Compound 1 can have a histologicallyor cytologically-confirmed IDH1 R132X gene-mutated advanced glioma thathas recurred or progressed following standard therapy. Patientsreceiving Compound 1 can have relapsed or refractory glioma (per WHOcriteria 2016) with confirmed IDH1 mutation. Other patients receivingCompound 1 can have relapsed or refractory hepatobiliary tumors withconfirmed IDH1 mutation previously treated with an approved therapy forHBC. Other patients receiving Compound 1 can have recurrent, refractoryor either locally advanced or metastatic chondrosarcoma with confirmedIDH1 mutation not amenable to complete surgical excision. Other patientsreceiving Compound 1 can have advanced, nonresectable or metastaticintrahepatic cholangiocarcinoma with confirmed IDH1 mutation noteligible for curative resection or transplantation. Patients can beassessed for pharmacokinetics (PK) (e.g., by collecting a blood sample)at regular intervals throughout a course of treatment. In particular,pre-dose PK assessment is performed at least on days 1, 2, 8, 15, and 22of the course of treatment for patients having a course of treatmentcomprising one or more 28-day treatment cycles. (Additional post doseassessment can be performed at cycle 1 day 1 and cycle 2 day 1.) Inaddition, pre-dose PK assessments are collected on day 1, 2 and 15 ofcycle 2 of a 28-day treatment cycle during the course of treatment.Additional pre-dose PK assessment is performed on day 1 of Cycle 3 andsubsequent 28-day treatment cycles during the course of treatment. Somepatients receiving Compound 1 can have relapsed or refractory othersolid tumors with confirmed IDH1 mutation.

In addition, methods of treatment can comprise the administration ofCompound 1 to patients who meet the following criteria for inclusion: 18years of age; Life expectancy of 4 months; Documented IDH1 gene-mutatedmalignancy based on local test evaluation; Able to provide tumor tissuesample (archival); and cancer diagnosis as detailed in below.Preferably, the methods comprise administering Compound 1 to a patientwho also meets one or more of the following additional inclusioncriteria:

-   -   Recovered to Grade 2 or baseline toxicity (except alopecia) from        prior therapy (per CTCAE v 4.03);    -   Eastern Cooperative Oncology Group (ECOG) performance status        0-2;    -   Adequate bone marrow function (e.g., Absolute neutrophil count        (ANC) ≥1.5×109/L without any growth factors in prior 7 days and        Hemoglobin 8.0 g/dL (with or without transfusion support);        Platelet count ≥75×10⁹/L (with or without transfusion support);        Cohort 4b (GemCis combination); and platelet count ≥100×109/L        (with or without transfusion support));    -   Child-Pugh Class A (HBC only)    -   Adequate hepatic function (e.g, Aspartate aminotransferase (AST)        (serum glutamic oxaloacetic transaminase [SGOT])/alanine        aminotransferase (ALT) (serum glutamic pyruvate transaminase        [SGPT]) ≤2.5× institutional upper limit of normal (ULN). For        patients with suspected malignancy related elevations, <5×ULN;        and Total bilirubin 1.5×ULN. For patients with suspected        malignancy related elevation <3× institutional upper limit of        normal); and    -   Adequate renal function (e.g, Creatinine clearance per        Cockcroft-Gault equation of ≥60 mL/min).

In some embodiments, Compound 1 is not administered to patients who meetone or more of the following exclusion criteria:

-   -   Previous solid organ or hematopoietic cell transplant    -   Prior anticancer treatment (e.g, No prior treatment with small        molecule, antibody, or other anticancer therapeutic within 21        days (or 5 half-lives), whichever is shorter of first dose of        study treatment (6 weeks for nitrosoureas or mitomycin C); No        previous treatment with an IDH1 inhibitor; No prior radiation        therapy (including radiofrequency ablation) within 4 weeks prior        to initiation of study treatment; No prior stereotactic body        radiation therapy (SBRT) within 2 weeks prior to initiation of        study treatment; and No prior chemoembolization or        radioembolization within 4 weeks)    -   Congestive heart failure (New York Heart Association Class III        or IV) or unstable angina pectoris; previous history of        myocardial infarction within one year prior to study entry,        uncontrolled hypertension, or uncontrolled arrhythmias    -   History of QT prolongation or baseline QT interval corrected        with Fridericia's method (QTcF) >450 ms (average of triplicate        readings) (NOTE: criterion does not apply to patients with a        right or left bundle branch block)    -   Concomitant medication(s) associated with QTc interval        prolongation or Torsades de Pointes (TdP) initiated less than        the duration required to reach steady-state plasma concentration        (approximately five half-lives) before first dose of study drug        (medications used as needed [PRN] (e.g. Zofran) are exempt).    -   Pregnant or nursing women or women of childbearing potential not        using adequate contraception; male patients not using adequate        contraception    -   Other malignancy within the last 5 years except: adequately        treated non-melanoma skin cancer, curatively treated in situ        cancer of the cervix, ductal carcinoma in situ (DCIS), stage 1,        grade 1 endometrial carcinoma, or other solid tumors including        lymphomas (without bone marrow involvement) curatively treated        with no evidence of disease for 5 years    -   Major surgery within 4 weeks of starting study treatment or not        recovered from any effects of prior major surgery (uncomplicated        central line placement or fine needle aspirate are not        considered major surgery);    -   Patients receiving >6 mg/day of dexamethasone or equivalent    -   Patients with gastrointestinal disorders likely to interfere        with absorption of the study medication    -   Known history of HIV positivity    -   Active infection with hepatitis B or C virus (Hep B or C viral        load>100 international units/milliliter or local institutional        equivalent)    -   Unstable or severe uncontrolled medical condition (e.g.,        unstable cardiac function, unstable pulmonary condition        including pneumonitis and/or interstitial lung disease,        uncontrolled diabetes) or any important medical or psychiatric        illness or abnormal laboratory finding that would, in the        Investigator's judgment, increase the risk to the patient        associated with his or her participation in the study and    -   PD-1 combination only: Patients with active autoimmune disease        (Note: patients with well controlled diabetes or hypothyroidism        are eligible).

Each patient receives Compound 1 throughout a medically appropriatecourse of treatment. In general, patients receive Compound 1 (eithersingle-agent Compound 1 or combination therapy as indicated above) untildisease progression or unacceptable toxicity. At the start of the courseof treatment, each patient receives Compound 1 as the solid formobtainable by the method of Example 1 at a dose of 150 mg BIDadministered continuously in one or more 28-day treatment cycles.

The DLT-Evaluable Analysis Set is defined as all patients in the SafetyLead-in Periods (single agent Compound 1, combination Compound1+5-azacitidine, combination Compound 1+GemCis and combination Compound1+PD-1 inhibitor such as nivolumab) who either experienced a DLT duringCycle 1 or completed at least 75% of the prescribed Cycle 1 dose. Thisanalysis sets will be used to assess the tolerability of Compound 1.

The Safety Analysis Set is defined as all patients who received anyamount of study drug(s) (Compound 1 and combination agents, ifappropriate).

This analysis set will be the primary analysis set for all safetyendpoints, excluding DLT evaluation. Safety analysis will be by cohortand by treatment within cohort if more than 1 dose or dosing combinationare initiated for a particular indication cohort.

The Response-Evaluable Analysis Set is defined as all patients withmeasurable disease at baseline who received the study drug(s) specificto the part of their particular cohort (either Compound 1 monotherapy orCompound 1 in combination), and had at least 1 post-baseline responseassessment or discontinued the treatment phase due to diseaseprogression (including death caused by disease progression) within 8weeks (+2-week window) of the first dose of study treatment. Thisanalysis set will be the primary analysis set for efficacy endpointssuch as ORR. All response evaluations will be by cohort, and bytreatment within cohort if more than 1 doses or dosing combinations areinitiated for a particular indication cohort.

Patient safety measurements and clinical laboratory measurements areperformed throughout the course of treatment for each cohort. Safetymeasurements include assessment of patient concomitant medications andprocedures, AE/SAE assessment, symptom-directed physical examination andECOG performance status. Clinical laboratory measurement assessmentincludes blood chemistry and hematology and other measurements specificto individual cohorts.

For patients receiving Compound 1 as a single agent, or in combinationwith 5-azacitidine (glioma, chondrosarcoma) or in combination with aPD-1 inhibitor (HBC), Compound 1 is administered in a 28-day treatmentcycle (28 consecutive days, at a dose of 150 mg BID) and patient safetymeasurement and clinical blood chemistry and hematology are obtained onthe following days during the course of treatment for patients in thefirst 28-day treatment cycle during the clinical trial: day 1, day 8(+/−2), day 15 (+/−2), day 22 (+/−2). In treatment cycle 2 and beyond,these assessments are obtained at day 1 (+/−2) and day 15 (+/−2).

For patients receiving Compound 1 in combination with chemotherapy(e.g., GemCis for cholangiocarcinoma), Compound 1 is administered in a28-day treatment cycle (28 consecutive days, at a dose of 150 mg BID)and patient safety measurement and clinical blood chemistry andhematology are obtained on the following days during the course oftreatment for patients in the first six 28-day treatment cycle duringthe clinical trial: day 1, day 8 (+/−2), day 15 (+/−2), day 22 (+/−2).The combination is given for a total of six treatment cycles. Intreatment cycle 7 and beyond, these assessments are obtained at day 1(+/−2) and day 15 (+/−2). The patient can continue on single agentCompound 1 treatment without combination agent as directed by treatingphysician.

Safety Lead-In Period

The study includes a Safety Lead-in Period where single-agent 150 mgCompound 1 BID administered over 28 days (1 cycle). The Safety-Lead-inPeriod employs a traditional 3+3 design, whereby 3 patients with solidtumors and 3 patients with gliomas are treated with Compound 1 150 mgBID and monitored for dose-limiting toxicities (DLTs) during the firstcycle of study treatment.

-   -   If no DLTs occur in the first 3 patients in either group (solid        tumors or glioma), and available pharmacokinetic        (PK)/pharmacodynamic (PD) data support the dose, enrollment        continues in the 4 disease-specific cohorts described below.    -   If a DLT occurs in the first 3 patients in either group, an        additional 3 patients are treated at that dose level in the        relevant group. If no DLTs occur in these additional 3 patients        (i.e. <2 DLTs per 6 patients) and available PK/PD data support        the dose, enrollment continues in the 4 disease-specific cohorts        described below.    -   If there are ≥2 DLTs at the starting dose, lower doses or an        altered dosing schedule of Compound 1 can be considered.        Likewise, higher doses may be evaluated based upon safety, PK,        and PD data as determined by the SRC.        Cohort 1: Glioma (n=16-31)

Compound 1 is used to treat patients diagnosed with a glioma cancerdiagnosis. In particular, Compound 1 is administered to patients meetingthe inclusion criteria above and one or more the following diseaserelated inclusion criteria: histologically or cytologically confirmedIDH1 gene-mutated advanced glioma, and a diagnosis of glioblastomamultiforme with confirmed IDH1 gene-mutated disease with first or secondrecurrence. Cohort 1 includes patients with glioma harboring an IDH1mutation that is relapsed or refractory. Glioma patients are treatedwith single-agent Compound 1 (Cohort 1a). Cohort 1a employs a Simon's2-stage design, in which 8 patients are treated with single-agentCompound 1 for a minimum of 4 cycles (cycle=28 days) and assessed forefficacy and safety (Stage 1). If 1 clinical response is observed inStage 1, then Stage 2 (n=15) initiates with single-agent Compound 1. Ifno clinical responses are observed in Stage 1 with single-agent Compound1, then combination therapy is examined (Compound 1+5-azacytidine)(Cohort 1b). Cohort 1b employs a Simon's 2-stage design, whereby 8patients are treated in Stage 1 with combination therapy for a minimumof 4 cycles (cycle=28 days) and assessed for efficacy and safety. If 1clinical response is observed in Stage 1 of Cohort 1b, then Stage 2(n=15) is initiated with combination therapy. During Stage 1 aggregatesafety data are monitored by the SRC. If unacceptable toxicity isobserved in Stage 1, then the dose and schedule may be modified by theSRC. (Note: any glioma patients enrolled in the safety Lead-in Periodare considered part of Stage 1 enrollment.)

Cohort 2: Hepatobiliary Carcinoma (HBC) (n=21-63)

Compound 1 is used to treat patients diagnosed with a hepatobiliarycarcinoma (HBC) cancer diagnosis. In particular, Compound 1 isadministered to patients meeting the inclusion criteria above and one ormore the following disease related inclusion criteria:Relapsed/refractory or intolerant to approved standard-of-care therapy(included: hepatocellular carcinoma, biliary carcinoma or otherhepatobiliary carcinomas); Histologically or cytologically confirmedIDH1 gene-mutated with measurable disease per RECIST 1.1 criteria; andChild-Pugh Class A.

Cohort 2 includes patients with relapsed/refractory HBC harboring anIDH1 mutation. HBC patients are initially treated with single-agentCompound 1 (Cohort 2a). Prior exposure to nivolumab is permitted forpatients of Cohort 2a. Cohort 2a employs a Simon's 2-stage design, inwhich 8 patients are treated with single-agent Compound 1 for a minimumof 4 cycles (cycle=28 days) and assessed for efficacy and safety (Stage1). If 1 clinical response is observed in Stage 1, then Stage 2 (n=15)is initiated with single-agent Compound 1. If no clinical responses areobserved in Stage 1 with single-agent Compound 1, then combinationtherapy can be examined (Compound 1+PD1 inhibitor) (Cohort 2b). Cohort2b employs a Simon's 2-stage design, whereby 13 patients are treated inStage 1 with combination therapy for a minimum of 4 cycles (cycle=28days) and assessed for efficacy and safety. If 4 clinical response isobserved in Stage 1 of Cohort 2b, then Stage 2 (n=42) can initiate withcombination therapy. Prior exposure to nivolumab is not permitted forpatients of Cohort 2b. During Stage 1 aggregate safety data is monitoredby the SRC. If unacceptable toxicity is observed in Stage 1, then thedose and schedule may be modified by the SRC. (Note: any HBC patientsenrolled in the Safety Lead-in Period are considered part of Stage 1enrollment.)

Cohort 3: Chondrosarcoma (n=16-31)

Compound 1 is used to treat patients diagnosed with a chondrosarcomacancer diagnosis. In particular, Compound 1 is administered to patientsmeeting the inclusion criteria above and one or more the followingdisease related inclusion criteria: Relapsed or refractory and eitherlocally advanced or metastatic and not amenable to complete surgicalexcision; and histologically or cytologically confirmed IDH1gene-mutated with measurable disease per RECIST 1.1 criteria. Cohort 3includes patients with relapsed/refractory, locally advanced ormetastatic chondrosarcoma harboring an IDH1 mutation. Chondrosarcomapatients are treated with single-agent Compound 1 (Cohort 3a). Cohort 3awill employ a Simon's 2-stage design, in which 8 patients will betreated with single-agent Compound 1 for a minimum of 4 cycles (cycle=28days) and assessed for efficacy and safety (Stage 1). If 1 clinicalresponse is observed in Stage 1, then Stage 2 (n=15) will initiate withsingle-agent Compound 1. If no clinical responses are observed in Stage1 with single-agent Compound 1, then combination therapy is examined(Compound 1+5-azacytidine) (Cohort 3b). Cohort 3b employs a Simon's2-stage design, whereby 8 patients are treated in Stage 1 withcombination therapy for a minimum of 4 cycles (cycle=28 days) andassessed for efficacy and safety. If 1 clinical response is observed inStage 1 of Cohort 3b, then Stage 2 (n=15) initiates with combinationtherapy. During Stage 1 aggregate safety data are monitored by the SRC.If unacceptable toxicity is observed in Stage 1, then the dose andschedule may be modified by the SRC. (Note: any chondrosarcoma patientsenrolled in the Safety Lead-in Period are considered part of Stage 1enrollment.)

Cohort 4: Intrahepatic Cholangiocarcinoma (n=21-63)

Compound 1 is used to treat patients diagnosed with an intrahepaticcholangiocarcinoma (IHCC) cancer diagnosis. In particular, Compound 1 isadministered to patients meeting the inclusion criteria above and one ormore of the following disease related inclusion criteria: Advancednonresectable or metastatic IHCC not eligible for curative resection ortransplantation; Phase 1b/Safety Lead-in of Phase 2: relapsed orrefractory disease; and histologically or cytologically confirmed IDH1gene-mutated with measurable disease per RECIST 1.1 criteria. Cohort 4includes patients with advanced IHCC harboring an IDH1 mutation. IHCCpatients are treated with single-agent Compound 1 (Cohort 4a). Patientsof cohort 4a must be ineligible for standard therapies. Cohort 4aemploys a Simon's 2-stage design, in which 8 patients are treated withsingle-agent Compound 1 for a minimum of 4 cycles (cycle=28 days) andassessed for efficacy and safety (Stage 1). If 2 clinical responses areobserved in Stage 1, then Stage 2 (n=15) initiates with single-agentCompound 1. If <2 clinical responses are observed in Stage 1 withsingle-agent Compound 1, then combination therapy is examined (Compound1+GemCis) (Cohort 4b). Patients in Cohort 4b have received no more thanone cycle of Gem/Cis therapy. Cohort 4b employs a Simon's 2-stagedesign, whereby 13 patients are treated in Stage 1 with combinationtherapy for a minimum of 4 cycles (cycle=21 days) and assessed forefficacy and safety. If 4 clinical responses are observed in Stage 1 ofCohort 4b, then Stage 2 (n=42) initiates with combination therapy.During Stage 1 aggregate safety data are monitored by the SRC. Ifunacceptable toxicity is observed in Stage 1, then the dose and schedulemay be modified by the SRC. (Note: any IHCC patients enrolled in theSafety Lead-in Period are considered part of Stage 1 enrollment.)

In some examples, patients diagnosed with relapsed/refractory IHCCreceive Compound 1 single agent, whereas patients newly diagnosed andtreatment naïve IHCC receive Compound 1 in combination with GemCischemotherapy.

Cohort 5: Other Non-CNS Solid Tumors with IDH1 Mutations (n=12)

Compound 1 is used to treat patients diagnosed with non-CNS solid tumorswith IDH1 mutations (preferably detectable 2-HG in plasma). Cohort 5includes patients with relapsed or refractory non-CNS solid tumorsharboring an IDH1 mutation. In particular, Compound 1 is administered topatients meeting the inclusion criteria above and one or more of thefollowing disease related inclusion criteria: relapsed or refractory tostandard-of-care therapy with no other available therapeutic options;and histologically or cytologically confirmed IDH1 gene-mutated withmeasurable disease per disease appropriate response criteria. Thiscohort includes treatment with single agent Compound 1. Due to thediverse population, this is an exploratory cohort without pre-definedefficacy/futility determinations. Aggregate safety data is monitored bythe SRC and if unacceptable toxicity is observed in 2 patients, thecohort can be closed for additional enrollment.

Table 25 Schedule of Efficacy Assessment by Cohort

For Cohorts 1-5 above, the efficacy assessments obtained include thoselisted in the table below. (MRS=1H magnetic resonance spectroscopy;MRI=magnetic resonance imaging).

TABLE 25 Assessment Response Cohort Population Criteria AssessmentBiomarkers Cycles (day 1) 1 Glioma Modified Contrast- N/A C3, C5, C7,C9, C12, RANO Criteria enhanced MRI every 3 cycles 2017, Low and MRS for2-HG thereafter Grade Glioma (central) RANO 2011 2 HBC Modified CT/MRISerum AFP, C3, C5, C7, C9, C12, RECIST CA19-9, every 3 cycles RECISTv1.1 and CEA thereafter 3 Chondro- RECIST v1.1 CT/18FDG-PET N/A C3, C5,C7, C9, C12, sarcoma every 3 cycles thereafter 4 Cholangio- RECIST v1.1CT/MRI Serum C3, C5, C7, C9, C12, carcinoma CA19-9, every 3 cycles(IHCC) and CEA thereafter 5 Other IDH1 Disease CT/MRI N/A C3, C5, C7,C9, C12, Solid appropriate every 3 cycles Tumors thereafter

We claim:
 1. A method of treating a patient diagnosed with acute myeloidleukemia (AML) having an IDH1 mutation, the method comprisingadministering to the patient in need thereof 150 mg of the compound ofFormula (1) twice per day (BID):


2. The method of claim 1, wherein the acute myeloid leukemia is relapsedor refractory or is drug-resistant.
 3. The method of claim 1, whereinthe compound of Formula (1) is orally administered to the patient. 4.The method of claim 1, further comprising administering 150 mg BID ofthe compound of Formula (1) to the patient throughout a course oftreatment of at least 15 consecutive days.
 5. The method of claim 4,further comprising administering 150 mg BID of the compound of Formula(1) to the patient on consecutive days throughout a course of treatmentof between 15 days and 6 months.
 6. The method of claim 4, furthercomprising administering 150 mg BID of the compound of Formula (1) tothe patient on consecutive days throughout a 6 month course oftreatment.
 7. The method of claim 4, wherein the compound of Formula (1)is administered to the patient in an oral unit dosage form.
 8. Themethod of claim 7, wherein the patient is diagnosed with acute myeloidleukemia that is relapsed or refractory or is drug-resistant prior tothe administration of the compound of Formula (1).
 9. A method oftreating a patient diagnosed with relapsed or refractory acute myeloidleukemia (AML) having an IDH1 mutation, the method comprising orallyadministering to the patient in need thereof a pharmaceuticalcomposition comprising 150 mg of the compound of Formula (1) twice perday (BID):

in an oral unit dosage form, on consecutive days throughout a course oftreatment of at least 15 consecutive days.
 10. The method of claim 9,further comprising administering to the patient in need thereof thecompound of Formula (1) on consecutive days for at least 6 months.